S-adenosylmethionine formulations with enhanced bioavailability

ABSTRACT

Provided herein are compositions and methods to enhance the absorption of S-adenosylmethionine (SAMe) and methods of treating various disorders or diseases using non-parenteral SAMe formulations with enhanced-absorption and improved bioavailability. In certain embodiments, the enhanced bioavailability formulations provided herein may be used to treat a variety of diseases or disorders, such as for example, psychiatric disorders including, generalized anxiety disorder, obsessive compulsive disorder, post traumatic stress disorder, panic disorder, depressive disorders (e.g. major clinical depression) and dysthymia; as well as treating liver disorders, cancer, autoimmune disorders, inflammatory disorders, joint disorders, gastrointestinal disorders and cardiovascular disease.

CROSS-REFERENCE TO RELATED APPLICATIONS AND CLAIM TO PRIORITY

This application claims priority to U.S. Non-Provisional patentapplication Ser. No. 12/845,555, filed Jul. 28, 2010; and claimspriority to U.S. Provisional patent application Ser. No. 61/435,997,filed Jan. 25, 2011 which are incorporated herein by reference in theirentirety.

TECHNICAL FIELD

Provided herein are compositions and methods for improvedbioavailability of 5-adenosyl-L-methionine (“SAM-e” or “SAMe”). Incertain embodiments, provided herein are formulations that modulateabsorption of exogenous SAMe throughout the gastrointestinal tract andthat provide, through oral administration or like method, a SAMe plasmaconcentration from which sufficient physiological effects can beexpected. Provided herein are methods of treating a disease or disorderin a subject and/or improving the nutritional status of a subject byadministering formulations enabling improved gastrointestinal absorptionof SAMe, wherein increased gastrointestinal absorption is achieved usingone or more absorption-enhancing technologies.

BACKGROUND OF THE INVENTION

S-adenosyl-L-methionine (“SAM-e” or “SAMe”) is a naturally occurringcompound that is present in tissues throughout the body. At themolecular level, SAMe is involved in various metabolic pathways,including transmethylation, transsulfuration and aminopropylation (e.g.in the production of polyamines, such as spermidine and spermine, fromputrescine).

S-adenosyl-L-methionine (SAMe)

In the body, SAMe is enzymatically synthesized from an amino acid,methionine, and a triphosphate nucleotide, ATP. SAMe has been tested innumerous clinical trials for the treatment of various ailments,including arthritis, liver disease and depression.

SAMe supplementation was initially considered impractical, due to theinstability of the SAMe ion during manufacturing, shipping and storage.Eventually stable salts of SAMe were developed (such as SAMe tosylatedisulfate, the butanedisulfonate salt of SAMe, the di-para-toluenesulfonate disulfate salt of SAMe, the tri-para-toluene sulfonic acidsalt of SAMe and the like). These salts can be formulated usingstandard, known technologies used for non-parenteral administrationincluding, but not limited to, tablets, capsules and pellets.Formulations such as these may also comprise a coating which can servemultiple purposes such as improving taste and ease of swallowing as wellas reducing stomach irritation. Stable salts of SAMe are described in,for example, U.S. Pat. Nos. 3,954,726 and 4,057,686, both of which areincorporated herein by reference in their entirety. Conventional SAMeActive Pharmaceutical Ingredient (API) is supplied as a molecular entitycomprising an ion along with several counter-ions. For example, SAMe ionplus a tosylate and 2 sulfonic acid counter-ions make up commerciallyavailable adenosylmethionine disulfate-p-toluenesulfonate (i.e. SAMetosylate disulfate). When referring to SAMe dosing, it is currentlyaccepted in the art that the numerical dose (usually in milligrams)refers to the amount of SAMe ion which is administered. For example,reference to a “400 mg SAMe tablet” using SAMe tosylate disulfate wouldinclude the 400 mg of SAMe ion, another 370 mg of the counter-ions, and200-300 mg of additional excipient to make up a final tablet weight of1.0-1.1 grams. Thus, for example, a 1600 mg oral dose of SAMe which isgenerally reported in the art would typically be a dose of four such1.0-1.1 gram tablets taken at one time. Alternatively, the same 1600 mgdose of SAMe ion may also be accomplished by administration of othercombinations of multiple tablets such as, sixteen 100 mg or eight 200 mgtablets of SAMe ion taken at a given time. Conventional oral dosageforms of SAMe are most commonly produced with about 400 mg of SAME ion;above that, the larger dosage form becomes difficult for swallowingconsidering that even at 400 mg of SAMe ion the tablets are quite largeat 1.0-1.1 grams.

The prevailing conventional wisdom in the art is of the view thatgastric juices in the stomach will alter the structure and/or functionof SAMe thereby reducing its absorption and therefore a pH-specificcoating which bypasses any SAMe release in the stomach is deemednecessary for oral administration. These “enteric” coatings are wellknown and routinely used by those of skill in the art as they arethought to provide a barrier which protects the encapsulated agent fromthe extremely low pH environment of the stomach. Although‘pH-sensitive’, these coatings are designed solely to protect theencapsulated agent from the stomach. They generally begin to dissolve ata pH above about 5.5 (designed to match the pH of the environmentimmediately following the stomach) to allow release of the underlyingdosage form. Various attempts to improve the stability and delivery ofenteric coated SAMe have been reported. Rao et al., describe the use ofan enteric coated, lipophilic soft gelatin capsule which beginsdissolving at pH 5.5 (U.S. Pat. No. 6,759,395). They recommend the useof a lipophilic material to insulate SAMe salts as a means of protectingthe encapsulated drug. Furthermore, they utilize a standard entericcoating in order to bypass any SAMe release in the stomach. The use ofan enteric coating is not surprising; in view of prior art reports thatSAMe cannot be absorbed in the stomach as it will be degraded first bygastric juices.

In addition to the reported claims in the art that SAMe is inactivatedwithin the stomach, there is also a widely accepted belief in these artssurrounding the absorption mechanism and metabolism of this compound.Based on past clinical experimentation, SAMe is cited as being highlysoluble and highly permeable yet exhibits low bioavailability. Studiesusing radiolabelled SAMe indicated that SAMe is readily absorbed withinthe gastrointestinal (GI) tract; however; plasma analysis showed verylow bioavailability (Stramentinoli, G., (1987) The American Journal ofMedicine 83(S 5A): 35-42). Therefore, those skilled in these artsassumed that SAMe's low bioavailability is caused by other factors, suchas “first pass metabolism” in the liver. Over the past 20 years,numerous groups have attempted to understand SAMe bioavailability bylooking at the pharmacokinetics, drug elimination and renal excretionprofiles of various SAMe formulations but not the absorption mechanisms.It is routinely reported by those most knowledgeable in these arts thatSAMe bioavailability when orally administered is limited to less than 5%because of “significant liver metabolism” prior to entering the blood(Bottiglieri et al., (1988) Alabama Journal of Medical Sciences 25(3):296-301; Bottiglieri et al., (1997) Exp. Opin. Invest. Drugs 6(4):417-426; Kaye et al., (1990) Drugs 40: 124-128). Additional drugelimination and renal excretion studies report that body accumulation ofintact SAMe is unlikely as a cause of reduced bioavailability andinstead also suggest that “active pre-systemic metabolism” is the cause(Giulidori and Cortellaro (1984) European Journal of ClinicalPharmacology 27: 119-121; Stramentinoli, G., (1986) BiologicalMethylation and Drug Design. R. T. Borchardt. New Jersey, Humana Press:315-326). Another belief is that metabolism of SAMe occurs rapidly viatransmethylation (and to a lesser extent, transsulfuration andaminoprophylation) pathways after non-parenteral administration. Morespecifically, the skilled practitioners of these arts proposed that themethyl group of SAMe is removed and incorporated into stable pools withlow turnover rates, such as proteins and phospholipids (Bottiglieri(1997) supra; Stramentinoli (1987) supra), and therefore results in thevery limited bioavailability of SAMe itself.

Active liver metabolism occurs with many drugs and typically causes alower cap on their bioavailability as seen with SAMe. Also, there is avast amount of clinical data reported in the art which supports readyabsorption of SAMe. It has thus been the general dogma in the art thatlow SAMe bioavailability is due primarily to extensive first passmetabolism in the liver.

A recent report looking at SAMe uptake into cells in culture finds thatSAMe is poorly transported through a monolayer of Caco-2 cells andpoorly absorbed by cultured rat hepatocytes (McMillan et al., (2005) J.of Pharmacy and Pharmacology, 57:599). There remains still a need toidentify both the reasons why exogenous SAMe bioavailability is low andways in which to increase it in vivo particularly for use in treatingdisease and/or nutritional status.

SUMMARY OF THE INVENTION

The present inventors have discovered that low permeability of SAMe isthe primary reason why: 1) in vivo SAMe bioavailability is limited, 2)SAMe exhibits different absorption patterns in different regions of theGI tract and, 3) levels of SAMe metabolites are not significantlyelevated after oral administration. Furthermore, the inventors also showfor the first time that it is possible in vivo to improve thebioavailability of SAMe by administering exogenous SAMe in combinationwith one or more absorption-enhancing technologies, especially tightjunction modulating agents. Provided herein are novel compositions andmethods that enhance the gastrointestinal absorption of SAMe.

The exemplary embodiments provided herein relate to methods andcompositions for enhancing the absorption of S-adenosyl-L-methionine(“SAMe”) or its stable salts as a means to increase SAMebioavailability. Use of compositions and methods provided herein in vivoimproves bioavailability of SAMe as compared to conventionalnon-parenteral dosage forms of SAMe which do not contain physiologicallyrelevant levels of absorption enhancers.

In certain embodiments, provided herein are non-parenteral compositionsof SAMe in combination with at least one absorption-enhancingtechnology. Without being limited to any particular theory,absorption-enhancing technologies which act to increase absorption of aphysiologically acceptable dosage of SAMe may work in a number of waysincluding, for example, increasing SAMe residence time in the GI tract(therefore allowing more opportunity for uptake); delivering SAMe toregions of the GI tract that exhibit increased drug absorption; adding“absorption enhancers” which increase either transcellular orparacellular transport of drugs (including agents which directly affecttight junction opening or penetration); encapsulating SAMe innanocarriers that deliver SAMe directly to cells; or a combination ofany of such technologies which modulate absorption. An“absorption-enhancing technology” is therefore any excipient, device,mechanism, technique, method, treatment parameter or the like whicheither directly or indirectly affects the absorption or uptake of SAMe.Many of these technologies may be designed to exploit or optimize SAMe'sinherent cationic nature at specific pH levels, for example, some mayact to maintain SAMe in its cationic form which is more easily absorbed(e.g. in the presence of a buffer or buffering system). Accordingly, incertain embodiments, the compositions provided herein can be combinedwith unconventional factors, such as diet (amount and/or type of foodand/or beverage), dosing schedule, the presence or absence of a coating(i.e. uncoated SAMe may be more efficiently absorbed) as a suitablemeans of altering SAMe absorption. In some embodiments, administrationof absorption-enhancing technologies prior to SAMe administration mayoptimize SAMe uptake.

Site-specific delivery of SAMe to segments of the GI tract exhibitingenhanced-absorption (also known as “absorption windows”) may be achievedwith the use of pH-dependent coatings which target SAMe release inpH-specific regions of the GI tract. As used herein, pH-dependentcoatings include any coating which bypasses release of SAMe in thestomach, for example: delayed release coatings, extended releasecoatings, enteric coatings and the like.

Some exemplary embodiments relate to compositions comprisingpH-dependent coatings, wherein the composition of the pH-dependentcoating acts to release a physiologically acceptable dosage of SAMe insegment-specific areas of the GI tract. In some embodiments,pH-dependent coatings allow release of SAMe in several regions along theentire GI tract in order to affect the site-specific effect of SAMeuptake and bioavailability. In some embodiments, absorption of SAMe mayoccur throughout the entire length of the GI tract. By identifyingregions with enhanced-absorption of SAMe, formulations targeted to theseregions can be administered to ensure better control of SAMe absorptionand bioavailability. In certain embodiments, pH-dependent coatings arenot employed as simple enteric coatings applied to avoid degradation inthe stomach. In certain embodiments, the pH-dependent coatings providedherein enable targeted delivery in the GI tract.

In some exemplary embodiments, pH-dependent coatings act to co-deliverSAMe and the absorption enhancer to the upper small intestine where SAMeis better absorbed. In other embodiments, the pH-dependent coatingresults in delivery of SAMe and the absorption enhancer to the largeintestine and/or colorectal regions, where SAMe is very poorly absorbed.A recent report in the art shows the inhibitory effects of SAMe on colonand gastric cancers cell growth in vitro (Luo et al., (2010) Int. J. ofBiological Sciences; 6(7); 784-795). The inventors discovered thatimproving absorption of SAMe in these areas, or targetingSAMe-containing dosage forms uncoating, release and dissolution in theseareas, particularly within the colorectal region, may provide a novelmeans for delivering SAMe within close proximity to these types oftumors as opposed to merely systemic delivery. Thus, in someembodiments, the pH-dependent coating is configured to deliver SAMedirectly to the colon and/or colorectal region, resulting in localand/or systemic delivery of SAMe to those parts of the intestinal tract.

Thus, certain exemplary embodiments also relate to methods forincreasing the bioavailability of SAMe by delivering pH-dependent coatedformulations of SAMe which act to release a physiologically acceptabledosage of SAMe and an absorption enhancer in site-specific ordisease-specific regions of the GI tract.

In certain embodiments, site-specific delivery of SAMe to segments ofthe GI tract exhibiting enhanced-absorption may be achieved with the useof non-pH-dependent coatings which target SAMe release in regions of theGI tract by delaying release of SAMe in a non-pH-dependent manner, e.g.for a period of time necessary for the dosage form to pass through theGI tract to the target portion of the GI tract, such as the duodenum,ileum, jejunum, colon and/or colorectal part of the GI tract.

In certain embodiments, several methods provided herein affect a delayedrelease with non-pH dependent polymers. These include soluble orerodible barrier systems, enzymatically degraded barrier systems,rupturable coating systems, and plugged capsule systems among others.Materials that can be used to obtain a delay in release suitable forthis component can be, but are not limited to, polyethylene glycol (PEG)with molecular weight above 4,000 daltons (Carbowax), PolyethyleneOxides (Polyox), waxes such as white wax or bees wax, paraffin, acrylicacid derivatives (Eudragit), propylene glycol, and ethylcellulose. Insome embodiments, these materials can be present in the range of 0.5-40%(W/W) of this component.

In certain embodiments, “absorption enhancers”, which also is meant toinclude agents known as “penetration enhancers”, “permeabilityenhancers” and “promoters” act directly on specific aspects of the GItract, such as paracellular transport, and affect the absorption rate ofnumerous drugs.

In certain embodiments, provided herein are compositions which make useof absorption enhancers to increase or promote absorption of aphysiologically acceptable dosage of SAMe as a mechanism for increasingSAMe bioavailability.

Certain exemplary embodiments relate to absorption enhancers whichdirectly modulate the activity of tight junctions. These are known astight junction penetration agents or tight junction modulating agents ortight junction opening agents. Tight junctions are intercellularjunctions between cells that control permeability between the cells. Inthis way, materials (e.g. APIs) cannot pass between cells but rathermust be taken up by the cell and thus enables the cells to regulate whatis allowed through. Tight junctions occur in many regions throughout thebody including the mouth, small intestine, large intestine and colon andvary in density/tightness within different regions. Within the GI tract,tight junctions refer to the areas between adjacent endothelial cellsand act to regulate the uptake of digested materials. Tight junctionsare highly regulated and are one of the key elements that form thebarrier between the luminal environment of the GI tract and the rest ofthe body.

Certain embodiments as exemplified herein also relate to compositionsand methods which incorporate tight junction opening agents to increaseor promote absorption of a physiologically acceptable dosage of SAMe.

Furthermore, the inventors show for the first time that, when studyingSAMe absorption, a subset of high potency tight junction openingabsorption enhancers exists and that identification of such high potencytight junction opening agents may be carried out using, for example, aCaco-2 cell model as described herein. Based on the concentration oftight junction modulator utilized as well as the increase in SAMepermeability achieved, suitable high potency tight junction modulatorselicit at least a two-fold increase in SAMe uptake in vitro when used atlow concentrations (i.e. about 2 mM or below). More preferably, at leasta three-fold increase in SAMe uptake occurs at low concentrations ofthese tight junction opening agents and, even more preferably, afour-fold or greater increase is achieved. Thus, some embodiments asexemplified herein also relate to compositions which incorporate highpotency tight junction opening agents to increase or promote absorptionof a physiologically acceptable dosage of SAMe. Certain otherembodiments as exemplified herein relate to compositions whichincorporate SAMe as well as one or more high potency tight junctionopening agents, wherein said tight junction opening agent yieldsenhanced absorption and causes minimal to no cell disruption as measuredin an in vitro cell permeability test. In certain embodiments, providedherein are methods for screening high potency tight junction openingagents. In some embodiments, a method provided herein is an in vitrocell permeability test. In some embodiments, a method provided herein isa method for screening high potency tight junction opening agent using aCaco-2 cell model.

In certain embodiments, pharmaceutical, medicinal, veterinary ornutritional preparations used for administering a physiologicallyacceptable dosage of SAMe include conventional solid or semi-solidtablets, pills, granules and capsules as well as controlled-releasetechnologies such as pH-sensitive drug targeting, timed-releasetechnologies, osmotic pumps, layered tablets, multiparticle tablets,nanocarriers or their combinations. When referring to “medicinal”preparations, purposes or treatments they are meant to include “medicalfoods”. Medical foods are defined by the U.S. Food and DrugAdministration as a food which is formulated to be consumed oradministered enterally under the supervision of a physician and which isintended for the specific dietary management of a disease or conditionfor which distinctive nutritional requirements, based on recognizedscientific principles, are established by medical evaluation.

Certain exemplary embodiments further relate to compositions fornon-parenteral administration of SAMe wherein SAMe is formulated in asolid or semi-solid composition which comprises one or moreabsorption-enhancing technology. In certain embodiments, provided hereinare methods of treatment wherein pharmaceutical, medicinal, veterinaryor nutritional preparations of SAMe are administered in conjunction withone or more absorption-enhancing technology. In a preferred embodiment,said absorption-enhancing technology is co-administered with saidpharmaceutical, medicinal, veterinary or nutritional preparations ofSAMe, and, even more preferably, said absorption-enhancing technology isincluded in said pharmaceutical, medicinal, veterinary or nutritionalpreparations of SAMe.

In certain embodiments, absorption-enhancing technologies need not formpart of the administered SAMe preparations and may be administeredseparately. Depending on their specific mechanism of action, the chosenabsorption-enhancing technology may be utilized either immediatelybefore, after or concurrent with the SAMe formulations. In someembodiments, provided herein are novel methods of treating a disease ordisorder in a subject in need thereof, wherein said method comprisesadministering a physiologically effective dosage of SAMe in combinationwith one or more absorption-enhancing technologies. In a preferredembodiment, said one or more absorption-enhancing technologies is atight junction modulating or opening agent.

Certain exemplary embodiments relate to methods for increasing thebioavailability of SAMe in a subject by delivering a composition fornon-parenteral administration comprising SAMe and at least oneabsorption-enhancing technology, wherein said absorption-enhancingtechnology acts either directly or indirectly to increase the absorptionof a physiologically acceptable dosage of SAMe.

In certain embodiments, diseases and/or disorders treatable with SAMeformulations provided herein are selected from the group consisting of,but not limited to, a mental or psychiatric disorder (e.g.psychotic/mood or non-psychotic mental disorders exemplified bydepression and substance related disorders, respectively), a nervoussystem disease/disorder (e.g. a central nervous system diseaseexemplified by Alzheimer's), other neurological disease/disorders (e.g.headaches and sleep disorders), conditions associated with injury to thecentral nervous system, a liver disease/disorder (e.g. alcoholic liverdisease), a cancer (e.g. solid and blood-borne cancers), a jointdisease/disorder (e.g. arthritis), an inflammatory disease/disorder(e.g. ulcerative colitis), an autoimmune disease/disorder (e.g. systemiclupus erythematosis and rheumatoid arthritis), a degenerativedisease/disorder (e.g. Amyotrophic Lateral Sclerosis), a soft-tissuedisease/disorder (e.g. a fibromyalgia disorder), a paindisease/disorder, a genetic disorder related to hyper- orhypo-methylation, a gastrointestinal disease/disorder, a cardiovasculardisease/disorder, a metabolic disease/disorder (e.g. Type 2 diabetes)and a disorder induced in whole or in part by oxidative or free-radicaldamage. Additional embodiments relate to combinations of SAMe with oneor more active ingredients that are commonly prescribed or used fortreatment of and/or prophylaxis of various diseases or disorders in asubject.

In certain embodiments, provided herein is a pharmaceutical compositioncomprising a therapeutically effective dosage of S-adenosylmethionineand at least one zwitterionic surfactant. In some embodiments, saidzwitterionic surfactant enhances systemic delivery ofS-adenosylmethionine as measured by an increase in C_(max) and/or AUCcompared to a control composition that lacks said zwitterionicsurfactant. In some embodiments, the C_(max) and/or AUC of saidcomposition is at least 140% of the C_(max) and/or AUC obtained with acontrol composition that lacks said zwitterionic surfactant.

In some embodiments, said composition is a non-parenteral composition.In some embodiments, the non-parenteral composition is an oral dosagecomposition, optionally formulated as a tablet, a capsule, or amultiparticulate formulation. In some embodiments, the non-parenteralcomposition is formulated as a dietary supplement or a medical food. Insome embodiments, further comprising at least one detergent, surfactant,zwitterionic surfactant, unsaturated cyclic urea, fatty acid, fattyamine, alkane sulfonate, bile acid, organic acid, cyclodextrin,chelating agent, salt of any of the foregoing, or a combination thereof.

In some embodiments, said zwitterionic surfactant is an acyl carnitine.In some embodiments, said acyl carnitine is a palmitoyl carnitine,lauroyl carnitine, stearoyl carnitine, myristoyl carnitine, decanoylcarnitine, or a salt thereof. In some embodiments, said zwitterionicsurfactant is a sulfobetaine. In some embodiments, said sulfobetaine issulfobetaine-10, sulfobetaine-12, sulfobetaine-14, sulfobetaine-16, orsulfobetaine-18.

In some embodiments, at least a portion of the composition is formulatedto dissolve in at least one of the stomach, duodenum, jejunum, ileum,large intestine, or colon. In some embodiments, the compositioncomprises a pH sensitive coating. In some embodiments, the compositionsprovided herein further comprise at least one delayed release component.In some embodiments, the delayed release component is a pH-triggeredenteric coating. In some embodiments, the delayed release component isformulated to dissolve in at least one of the duodenum, jejunum, ileum,large intestine, or colon.

In some embodiments, S-adenosylmethionine and said zwitterionicsurfactant are in a ratio of from 1:100,000 to 100,000:1, or 1:150 to150:1, or 1:10 to 10:1 (weight:weight). In some embodiments,S-adenosylmethionine and said zwitterionic surfactant are in a ratio of1:1 (weight:weight). In some embodiments, the composition comprises 50to 3200 mg of S-adenosylmethionine. In some embodiments, the compositioncomprises 50 to 1600 mg of S-adenosylmethionine.

In some embodiments, the compositions provided herein further compriseat least one delayed release component. In some embodiments, the delayedrelease component is a pH-triggered enteric coating. In someembodiments, the delayed release component is formulated to dissolve inat least one of the duodenum, jejunum, ileum, large intestine, or colon.

In certain embodiments, provided herein are compositions for use in thetreatment of a condition selected from the group consisting of a mentalor psychiatric disorder, a nervous system disease or disorder, aneurological disease or disorders, a condition associated with injury tothe central nervous system, a liver disease or disorder, a cancer, ajoint disease or disorder, an inflammatory disease or disorder, anautoimmune disease or disorder, a degenerative disease or disorder, asoft-tissue disease or disorder, a pain disease or disorder, a geneticdisorder related to hyper- or hypo-methylation, a gastrointestinaldisease or disorder, a cardiovascular disease or disorder, and adisorder induced in whole or in part by oxidative or free-radicaldamage.

In certain embodiments, provided herein are methods for increasing thebioavailability of exogenous SAMe administered to a subject, said methodcomprising administering to the subject a non-parenteral compositioncomprising a therapeutically effective dosage of S-adenosylmethionineand at least one zwitterionic surfactant. In some embodiments, thecomposition is an oral dosage composition. In some embodiments, thecomposition is formulated as a dietary supplement or a medicinal food.In some embodiments, the compositions further comprise a detergent, asurfactant, a zwitterionic surfactant, an unsaturated cyclic urea, afatty acid, a fatty amine, an alkane sulfonate, a bile acid, an organicacid, a cyclodextrin, a chelating agent, a salt of any of the foregoing,or a combination thereof. In some embodiments, at least a portion of thecomposition is formulated to dissolve in at least one of the stomach,duodenum, jejunum, ileum, large intestine, or colon. In someembodiments, the composition comprises a pH sensitive coating. In someembodiments, said zwitterionic surfactant is administered either beforeor after administration of the composition comprising the at least onephysiologically effective dosage of S-adenosylmethionine.

In certain embodiments, provided herein are methods of treating adisorder selected from the group consisting of a mental or psychiatricdisorder, a nervous system disease or disorder, a neurological diseaseor disorders, a condition associated with injury to the central nervoussystem, a liver disease or disorder, a cancer, a joint disease ordisorder, an inflammatory disease or disorder, an autoimmune disease ordisorder, a degenerative disease or disorder, a soft-tissue disease ordisorder, a pain disease or disorder, a genetic disorder related tohyper- or hypo-methylation, a gastrointestinal disease or disorder, acardiovascular disease or disorder, and a disorder induced in whole orin part by oxidative or free-radical damage, comprising administering toa patient in need thereof a composition provided herein.

In certain embodiments, provided herein is a non-parenteral compositioncomprising at least one physiologically effective dosage ofS-adenosylmethionine in combination with at least oneabsorption-enhancing technology. In some embodiments, theabsorption-enhancing technology is one of gastroretentive dosageadjuvants, gastrointestinal segment-specific delivery systems,chemically derived absorption enhancing agents, tight junctionpenetration agents, tight junction opening agents, nanocarriers, a dietregimen, and a dosing regimen. In some embodiments, the non-parenteralcomposition is an oral dosage composition. In some embodiments, thenon-parenteral composition is incorporated in a dietary supplement or amedical food.

In some embodiments, the non-parenteral dosage composition comprises atleast one physiologically effective dosage of S-adenosylmethionine incombination with at least one of a tight junction penetration agent andtight junction opening agent. In some embodiments, the at least one of atight junction penetration agent and tight junction opening agent isselected from the group consisting of detergents, surfactants,zwitterionic surfactants, unsaturated cyclic ureas, fatty acids, fattyamines, alkane sulfonates, bile acids, organic acids, cyclodextrins,chelating agents, salts of any of the foregoing, and combinationsthereof. In some embodiments, at least one of a tight junctionpenetrating agent and a tight junction opening agent includes azwitterionic surfactant. In some embodiments, the zwitterionicsurfactant is an acyl carnitine. In some embodiments, the zwitterionicsurfactant is a sulfobetaine. In some embodiments, at least one of atight junction penetrating agent and a tight junction opening agentincludes a fatty acid or a salt thereof. In some embodiments, at leastone of a tight junction penetrating agent and a tight junction openingagent includes a fatty amine or a salt thereof. In some embodiments, atleast one of a tight junction penetrating agent and a tight junctionopening agent includes a bile acid or a salt thereof. In someembodiments, at least one of a tight junction penetrating agent and atight junction opening agent includes detergent, a surfactant, anunsaturated cyclic urea, and organic acid, a cyclodextrin, a chelatingagent, a salt of any thereof, or a combination of two or more thereof.

In some embodiments, at least a portion of the composition is configuredto dissolve in at least one of the stomach, duodenum, jejunum and ileum.In some embodiments, at least a portion of the composition is configuredto dissolve in the large intestine or colon. In some embodiments, thecomposition incorporates a pH sensitive coating.

In certain embodiments, provided herein is a method for increasing thebioavailability of exogenous SAMe administered to a subject, said methodcomprising administering to the subject a non-parenteral compositioncomprising at least one physiologically effective dosage ofS-adenosylmethionine in combination with at least oneabsorption-enhancing technology. In some embodiments, theabsorption-enhancing technology is one of gastroretentive dosageadjuvants, gastrointestinal segment-specific delivery systems,chemically derived absorption enhancing agents, tight junctionpenetration agents, tight junction opening agents, nanocarriers, a dietregimen, and a dosing regimen. In some embodiments, the composition isan oral dosage composition. In some embodiments, the composition isincorporated in a dietary supplement or a medicinal food. In someembodiments, the composition comprises a physiologically effectivedosage of S-adenosylmethionine in combination with at least one of atight junction penetration agent and tight junction opening agent. Insome embodiments, the composition comprises at least one of a tightjunction penetration agent and tight junction opening agent is selectedfrom the group consisting of detergents, surfactants, zwitterionicsurfactants, unsaturated cyclic ureas, fatty acids, fatty amines, alkanesulfonates, bile acids, organic acids, cyclodextrins, chelating agents,salts of any of the foregoing, and combinations thereof. In someembodiments, the composition comprises at least one of a tight junctionpenetrating agent and a tight junction opening agent includes azwitterionic surfactant. In some embodiments, the composition comprisesat least one of a tight junction penetrating agent and a tight junctionopening agent includes a fatty acid or a salt thereof. In someembodiments, at least one of a tight junction penetrating agent and atight junction opening agent includes a fatty amine or a salt thereof.In some embodiments, at least one of a tight junction penetrating agentand a tight junction opening agent includes a bile acid or a saltthereof. In some embodiments, at least one of a tight junctionpenetrating agent and a tight junction opening agent includes detergent,a surfactant, an unsaturated cyclic urea, and organic acid, acyclodextrin, a chelating agent, a salt of any thereof, or a combinationof two or more thereof. In some embodiments, at least a portion of thecomposition is configured to dissolve in at least one of the stomach,duodenum, jejunum and ileum. In some embodiments, at least a portion ofthe composition is configured to dissolve in the large intestine orcolon. In some embodiments, the composition incorporates a pH sensitivecoating. In some embodiments, the absorption-enhancing technology isadministered either before or after administration of the compositioncomprising the at least one physiologically effective dosage ofS-adenosylmethionine.

In some embodiments, provided herein is a method of treating in apatient a disorder selected from the group consisting of a mental orpsychiatric disorder (e.g. psychotic/mood or non-psychotic mentaldisorders exemplified by depression and substance related disorders,respectively), a nervous system disease/disorder (e.g. a central nervoussystem disease exemplified by Alzheimer's), other neurologicaldisease/disorders (e.g. headaches and sleep disorders), conditionsassociated with injury to the central nervous system, a liverdisease/disorder (e.g. alcoholic liver disease), a cancer (e.g. solidand blood-borne cancers), a joint disease/disorder (e.g. arthritis), aninflammatory disease/disorder (e.g. ulcerative colitis), an autoimmunedisease/disorder (e.g. systemic lupus erythematosis and rheumatoidarthritis), a degenerative disease/disorder (e.g. Amyotrophic LateralSclerosis), a soft-tissue disease/disorder (e.g. a fibromyalgiadisorder), a pain disease/disorder, a genetic disorder related to hyper-or hypo-methylation, a gastrointestinal disease/disorder, acardiovascular disease/disorder, and a disorder induced in whole or inpart by oxidative or free-radical damage, comprising administering tothe patient in need thereof a composition provided herein.

In some embodiments, provided herein is a solid oral compositioncontaining a physiologically effective amount of S-adenosylmethionineand a tight junction opening absorption enhancer at a ratio of from1:100,000 to 100,000:1 (weight:weight). In some embodiments, theS-adenosylmethionine and the absorption enhancer are in a ratio of from1:150 to 150:1. In some embodiments, the 5-adenosylmethionine and theabsorption enhancer are in a ratio of from 1:10 to 10:1. In someembodiments, the solid oral composition further comprises at least onedelayed release component. In some embodiments, the delayed releasecomponent is a pH-triggered, enteric coating applied to the sold oralcomposition. In some embodiments, the delayed release component is suchthat the composition is configured to dissolve in at least one of theduodenum, jejunum, and ileum. In some embodiments, the delayed releasecomponent is such that the release is configured to dissolve in thelarge intestine or colon. In some embodiments, the tight junctionopening absorption enhancer is a high potency absorption enhancer asdetermined in a Caco-2 monolayer permeability assay. In someembodiments, the tight junction opening absorption enhancer comprisesone or more detergents, surfactants, zwitterionic surfactants,unsaturated cyclic ureas, fatty acids, fatty amines, alkane sulfonates,bile acids, organic acids, cyclodetrins, chelating agents, andcombinations thereof. In some embodiments, the tight junction openingabsorption enhancer comprises one or more zwitterionic surfactants. Insome embodiments, the one or more zwitterionic surfactants includes asulfobetaine. In some embodiments, the tight junction opening absorptionenhancer includes a fatty amine or a salt thereof. In some embodiments,the tight junction opening absorption enhancer includes a bile acid or asalt thereof. In some embodiments, the tight junction opening absorptionenhancer includes a detergent, a surfactant, an unsaturated cyclic urea,an organic acid, a cyclodextrin, a chelating agent, a salt of anythereof, or a combination of two or more thereof. In some embodiments,the tight junction opening absorption enhancer includes a long-chainacyl carnitine zwitterionic surfactant or a salt thereof. In someembodiments, the tight junction opening absorption enhancer includes oneof the long-chain acyl carnitine zwitterionic surfactants selected frompalmitoyl carnitine, lauroyl carnitine, stearoyl carnitine, myristoylcarnitine, and decanoyl carnitine or a salt thereof. In someembodiments, the tight junction opening absorption enhancer includespalmitoyl carnitine or a salt thereof. In some embodiments, thecomposition is in an oral dosage form. In some embodiments, the oraldosage form is a tablet, a capsule or a multiparticulate formulation. Insome embodiments, the dosage form comprises about 50 to about 500 mg ofS-adenosylmethionine.

In some embodiments, provided herein is a composition for the treatmentof a condition amenable to treatment with S-adenosylmethionine.

In some embodiments, provided herein is a method of treating a conditionamenable to treatment with S-adenosylmethionine, comprisingadministering to a patient a physiologically effective amount of acomposition provided herein.

In some embodiments, provided herein is a pharmaceutical, solid oralcomposition containing a physiologically effective dose ofS-adenosylmethionine and a tight junction opening absorption enhancer,wherein the enhancer enhances systemic delivery of S-adenosylmethionineas measured by an increase in Cmax and/or AUC compared to a controlcomposition that lacks said enhancer. In some embodiments, the Cmaxand/or AUC obtained with the dose of S-adenosylmethionine and a tightjunction is at least 140% of the Cmax and/or AUC obtained with thecontrol formulation or greater. In some embodiments, solid oralcomposition further comprises at least one delayed release component. Insome embodiments, the delayed release component is a pH-triggered,enteric coating applied to the sold oral composition. In someembodiments, the delayed release component is such that the compositionis configured to dissolve in at least one of the duodenum, jejunum, andileum. In some embodiments, the delayed release component is such thatthe release is configured to dissolve in the large intestine or colon.In some embodiments, the tight junction opening absorption enhancer is ahigh potency absorption enhancer as determined in a Caco-2 monolayerpermeability assay. In some embodiments, the tight junction openingabsorption enhancer comprises one or more detergents, surfactants,zwitterionic surfactants, unsaturated cyclic ureas, fatty acids, fattyamines, alkane sulfonates, bile acids, organic acids, cyclodetrins,chelating agents, and combinations thereof. In some embodiments, thetight junction opening absorption enhancer comprises one or morezwitterionic surfactants. In some embodiments, the one or morezwitterionic surfactants includes a sulfobetaine. In some embodiments,the tight junction opening absorption enhancer includes a fatty amine ora salt thereof. In some embodiments, the tight junction openingabsorption enhancer includes a bile acid or a salt thereof. In someembodiments, the tight junction opening absorption enhancer includes adetergent, a surfactant, an unsaturated cyclic urea, an organic acid, acyclodextrin, a chelating agent, a salt of any thereof, or a combinationof two or more thereof. In some embodiments, the tight junction openingabsorption enhancer includes a long-chain acyl carnitine zwitterionicsurfactant or a salt thereof. In some embodiments, the tight junctionopening absorption enhancer includes one of the long-chain acylcarnitine zwitterionic surfactants selected from palmitoyl carnitine,lauroyl carnitine, stearoyl carnitine, myristoyl carnitine, and decanoylcarnitine or a salt thereof. In some embodiments, the tight junctionopening absorption enhancer includes palmitoyl carnitine or a saltthereof. In some embodiments, the composition is in an oral dosage form.In some embodiments, the oral dosage form is a tablet, a capsule or amultiparticulate formulation. In some embodiments, the dosage formcomprises about 50 to about 500 mg of S-adenosylmethionine.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A is a full-scale graph of the average plasma concentration ofSAMe versus time of subjects who were administered 800 mg of one ofthree segment-specific SAMe formulations comprising coatings designed torelease SAMe in the proximal GI tract (duodenum/jejunum; squares), inthe distal GI tract (ileum/ascending colon; triangles) and meteredthroughout the entire GI tract (circles); and

FIG. 1B is a magnified view of the graph in FIG. 1A which betterhighlights the separation between the lower average plasma concentrationcurves;

FIG. 2 is a graph showing the permeability of SAMe across a monolayer ofCaco-2 human colonic adenocarcinoma cells alone and in the presence ofEDTA or in the absence of calcium. Propranolol is included as a highpermeability control;

FIG. 3A is a graph showing the permeability of SAMe across a monolayerof Caco-2 human colonic adenocarcinoma cells alone and in the presenceof various tight junction modulators; and

FIG. 3B is a graph showing the average plasma concentration of SAMeversus time when SAMe is delivered both in the presence and absence ofthe tight junction modulating agent, caprate. Animal subjects wereadministered either a single dose of SAMe disulfate tosylate incombination with inactive excipients and sodium caprate formulated in anenteric-coated capsule, or the control consisting of the identicalformulation but without sodium caprate; and

FIG. 3C is a graph showing the average plasma concentration of SAMeversus time when SAMe is delivered both in the presence and absence ofthe tight junction modulating agent, palmitoyl-DL-carnitine chloride(PCC). Animal subjects were administered either a single dose of SAMedisulfate tosylate in combination with inactive excipients and PCCformulated in an enteric-coated capsule, or the control consisting ofthe identical formulation but without PCC.

FIG. 4A is a graph showing the average plasma concentration of SAMeversus time when SAMe is delivered both in the presence and absence ofthe tight junction modulating agent, Sulfobetaine 16. Animal subjectswere administered either a single dose of SAMe disulfate tosylate incombination with inactive excipients and Sulfobetaine 16 formulated at a1:1 ratio (w/w) in an enteric-coated capsule, or the control consistingof the identical formulation but without Sulfobetaine 16; and

FIG. 4B is a graph showing the average plasma concentration of SAMeversus time when SAMe is delivered both in the presence and absence ofthe tight junction modulating agent, Sulfobetaine 14. Animal subjectswere administered either a single dose of SAMe disulfate tosylate incombination with inactive excipients and Sulfobetaine 14 formulated at a1:1 ratio (w/w) in an enteric-coated capsule, or the control consistingof the identical formulation but without Sulfobetaine 14.

FIG. 5 is a graph of the average plasma concentration of SAMe as well asthe SAMe metabolite, S-adenosyl homocysteine (SAH), versus time fromseven subjects administered a 1600 mg dose of commercially availableSAMe tosylate disulfate;

FIG. 6 is a graph of the average plasma concentration of SAMe versustime from seven subjects each administered a 400 mg dose of an uncoatedoral formulation of SAMe.

DETAILED DESCRIPTION OF THE INVENTION

The inventors have discovered that contrary to the general state of theart, SAMe bioavailability is low because it is poorly absorbed and alsothat SAMe demonstrates distinct absorption patterns within differentregions of the GI tract in humans. Furthermore, they found that thelevel of SAMe metabolites in the blood are minimally affected uponexogenous SAMe administration, which also clearly suggests that lowbioavailability of exogenous SAMe is not primarily due to extensivefirst pass metabolism in the liver. Finally, known tight junctionmodulators significantly increase the permeability of SAMe across amodel monolayer of cells and in vivo delivery of certain tight junctionopening absorption enhancers in combination with exogenous SAMe resultin substantial increases in SAMe bioavailability. The importance ofthese discoveries is significant and several techniques are providedherein which can alter and increase the gastrointestinal absorption ofSAMe.

The inventors discovered that because SAMe permeability is low, it ispossible to increase SAMe bioavailability in vivo by utilizing factorswhich enhance the absorption rate of this compound.

Some exemplary embodiments relate to compositions that modulate andimprove the absorption and bioavailability of non-parenterallyadministered SAMe. Related exemplary embodiments provide methods ofusing the compositions for therapeutic treatment of certain diseasesand/or disorders and/or as nutritional supplements and/or as medicalfoods. Additional embodiments relate to combinations of SAMe with one ormore active ingredients that are commonly prescribed or used fortreatment of and/or prophylaxis of various diseases or disorders in asubject.

As used herein the term “SAMe” refers to S-adenosyl-L-methionine and itsvariant, S-adenosylmethionine. As shown in the structural formulapresented earlier, SAMe appears as a charged species, and its ionizationstate varies with pH. As mentioned previously, in its solid form, SAMeis present as a salt comprised of the SAMe ion as well as one or morecounter-ions. It is common to find SAMe in a stable salt form (e.g. withp-toluenesulfonic acid as the negative counter ion) alone or incombination with one or more additional salt-forming substances, forexample, mineral or organic acids and/or amino acids (See U.S. Pat. No.3,893,999, incorporated herein by reference in its entirety). Otherstable SAMe salts are described in, for example, U.S. Pat. No.5,128,249, which discloses particular stable salts of SAMe. Variousmorphologies of SAMe are suitable for use in the present embodiments.Thus, as used herein “SAMe” refers to the stable salts and amorphousforms and semicrystalline forms and crystalline forms of SAMe as well asto the ionic form of SAMe when present in vivo. Amorphous forms of SAMecan be employed at any particle size and particle size distribution.

In certain embodiments, formulations for non-parenteral administrationof SAMe are provided as solid or semi-solid products or dosage forms,such as tablets, capsules or pellets, and generally consist of a core“matrix material” which ‘encapsulates’ the drug as well as one or moreprotective coatings. “Product” or “dosage form” as used herein refers toany solid or semi-solid formulation or preparation used for non-parentaladministration of SAMe. In certain embodiments, non-parenteralformulations or preparations as described herein include oral deliverysystems exemplified by tablets, pastes, capsules, granules, caplets,lozenges and the like; and transdermal, transmucosal or inhaled deliverysystems, exemplified by aerosols, irrigants, topical creams, pastes,patches, lozenges and the like, all of which are well-known andwell-documented in the art. In some embodiments, these formulations maybe administered using a clinical, pharmaceutical or veterinary dosingregimen. In some embodiments, non-parenteral SAMe dosage forms may alsobe provided as medical foods or dietary or nutritional supplements.

In certain embodiments, non-parenterally administered SAMe formulationsmay be configured to enable extended release of the encapsulated SAMe.Co-owned U.S. patent application 2009/0088404, which is incorporatedherein by reference, provides novel formulations of extended-releaseSAMe formulations. As disclosed in U.S. 2009/0088404, there are avariety of methods which can be used to prepare extended-releasecompositions of various types of drugs; and it is contemplated that atleast one of these methodologies can be used to prepare extended-releaseSAMe compositions with enhanced bioavailability properties. The types ofextended-release SAMe compositions that are contemplated within thescope of the present embodiments include osmotic dosage forms,extended-release matrices, pulsatile-release formulations andextended-release formulations coated with one or more enteric coatingsall of which are described in detail in U.S. 2009/0088404.

A “physiologically effective dosage” of SAMe as used herein is meant toinclude an amount of SAMe which is administered under a defined dosingregimen for either clinical, pharmaceutical, medicinal, veterinary,dietary or nutritional purposes. Thus a “physiologically effectivedosage” of SAMe includes a therapeutically effective dosage, apharmaceutically acceptable dosage, a veterinary acceptable dosage, anutraceutically acceptable dosage, a dietary acceptable dosage and anutritionally acceptable dosage of SAMe as well as an acceptable dosagefor use as a medical food and all of which are included for use in thepresent embodiments.

In certain embodiments, the relative bioavailability of SAMeformulations is determined by assessing its pharmacokinetic profileusing well known techniques such as area under the curve (AUC; which isa measure of the overall exposure of a subject to SAMe in the plasmaafter a dose), Cmax (i.e. the highest concentration of SAMe in theplasma that is measured after a dose and T_(max) (i.e. the time afteradministration of a drug when the maximum plasma SAMe concentration isreached)—all of these measurements are extensively described in the art.

In some embodiments, provided herein is a method for treating and/orprophylaxis in a subject a disorder selected from the group consistingof, but not limited to, a mental or psychiatric disorder (e.g.psychotic/mood or non-psychotic mental disorders exemplified bydepression and substance related disorders, respectively), a nervoussystem disease/disorder (e.g. a central nervous system diseaseexemplified by Alzheimer's), other neurological disease/disorders (e.g.headaches and sleep disorders), conditions associated with injury to thecentral nervous system, a liver disease/disorder (e.g. alcoholic liverdisease), a cancer (e.g. solid and blood-borne cancers), a jointdisease/disorder (e.g. arthritis), an inflammatory disease/disorder(e.g. ulcerative colitis), an autoimmune disease/disorder (e.g. systemiclupus erythematosis and rheumatoid arthritis), a degenerativedisease/disorder (e.g. Amyotrophic Lateral Sclerosis), a soft-tissuedisease/disorder (e.g. a fibromyalgia disorder), a paindisease/disorder, a genetic disorder related to hyper- orhypo-methylation, a gastrointestinal disease/disorder, a cardiovasculardisease/disorder, a metabolic disease/disorder (e.g. Type 2 diabetes)and a disorder induced in whole or in part by oxidative or free-radicaldamage, comprising administering to said subject an exemplarycomposition provided herein which enhances the absorption of aphysiologically effective dosage of SAMe, whereby theenhanced-absorption provides an increase in SAMe bioavailability.

Some exemplary embodiments relate to compositions and methods of theiruse for enhancing the effectiveness of a physiologically effectivedosage of SAMe utilized as a dietary or nutritional supplement in asubject. In some embodiments, effectiveness as a dietary or nutritionalsupplement may be measured using one or more nutritional performancevariables, such as improved concentration, memory, mood, nutritionalstatus or liver status.

Absorption-Enhancing Technologies as a Means of Improving SAMeAbsorption and Bioavailability

Through the discovery that SAMe absorption is a limiting factor in thesystemic bioavailability of SAMe, provided herein are means ofincreasing or modulating its absorption. Any method which eitherdirectly or indirectly enhances SAMe absorption throughout the body iscontemplated within the scope of the present embodiments, including forexample, increasing SAMe residence time in the GI tract thereby allowingmore opportunity for uptake, delivering SAMe to targeted regions of theGI tract that exhibit increased drug absorption characteristics,incorporation of “absorption enhancers” (including “penetrationenhancers” and “promoters”) which increase either transcellular and/orparacellular transport of drugs (including agents which directly affecttight junctions); encapsulating SAMe in nanocarriers that deliver SAMedirectly to cells; maintaining SAMe in its cationic form, modulatingdiet and/or dosing schedule, delivering SAMe uncoated or a combinationof any of such ‘technologies’ which modulate absorption. When referringto the “gastrointestinal tract” or “GI tract”, it is intended to includethe entire region beginning with the mouth/cheeks through to theesophagus, stomach, small intestine, large intestine and colorectalregions.

Mechanisms to Increase Gastric Retention Time

In certain embodiments, increasing SAMe gastric retention time may beachieved using, for example, gastroretentive dosage forms (GRDF) of thedrug including floating, geometric, bioadhesive and swelling dosageforms which are designed to withstand peristalsis and mechanicalcontractility of the stomach.

GI Segment-Specific Targeted Formulations

In certain embodiments, site-specific delivery of SAMe to multiple sitesalong the GI tract is useful in understanding and modulating SAMeabsorption since the unique environment of different segments throughoutthe intestinal tract can affect absorption of different drugs. Inparticular, drugs which show low permeability in the GI tract tend to beabsorbed in specific areas along the tract. Thus, in some embodiments,their delivery site may be controlled in order to control theabsorption.

In certain embodiments, targeted delivery sites in the GI tract includeone or more of the mouth, stomach, duodenum, jejunum, ileum, colon andrectum. The pH along the GI tract varies from as low as 1 in the stomachto 8 in certain segments of the intestines. The GI tract is a highlycomplex environment with distinct pH zones that vary in locationdepending on a number of factors, including diet. Typically the pHranges from lowest in the stomach to higher pH zones in the small andlarge intestine.

The large intestine is the final organ comprising the GI tract andincludes the colon and rectum. The large intestine is the site for waterresorption and formation of feces. Like the buccal area, blood thatdrains the rectum is not first transported to the liver. Therefore,absorption that takes place in the rectum (e.g., from rectalsuppositories and enemas) enters the systemic circulation system withoutany biotransformation that may otherwise have occurred in the liver.

In addition to pH and/or tight junction porosity, other physiologicalfactors such as surface area, enzymatic and transporter activity, aswell as colonic microflora influence drug absorption, and it is withinthe scope of the present embodiments to modulate one or more of thesefactors in any region(s) of the GI tract as a means of affecting thebioavailability of SAMe.

Paracellular transport, mediated through tight junctions is higher inthe proximal segments of the GI tract, exemplified by the duodenum,jejunum and ileum. Paracellular transport is much less in the distalsegments, such as the colon.

Some exemplary embodiments relate to novel compositions comprisingpH-dependent coated SAMe in combination with at least one absorptionenhancing agent, wherein the composition of the pH-dependent coatingacts to release a physiologically acceptable dosage of SAMe insegment-specific areas of the GI tract. In some embodiments,pH-dependent coatings may be configured to enable release of SAMe inseveral regions along the entire GI tract in order to affectsite-specific absorption and bioavailability of SAMe.

Some exemplary embodiments relate to compositions comprising SAMe innon-enteric coated (or “uncoated”) formulations. In contrast to thecurrent general state of the art, the inventors found that SAMe can beeffectively released into the stomach and give rise to modestly elevatedSAMe plasma levels and therefore an enteric coating is not critical forpreventing degradation.

Absorption Enhancers

The epithelial and endothelial barriers of the human body provide majorobstacles for drug delivery to the systemic circulation systems and alsoto organs with unique environments, such as the central nervous system.Several transport routes exist in these barriers, which potentially canbe exploited for enhancing drug permeability and absorption. Compared tothe transcellular pathways (via transporters, adsorptive andreceptor-mediated transcytosis), the paracellular flux for cells andmolecules is very limited. Over the past 40 years many groups have beendeveloping absorption or permeability enhancers. These “promoters” aregenerated as a means of modifying intercellular junctions andparacellular permeability.

Thus, some exemplary embodiments relate to compositions comprising aphysiologically acceptable dosage of SAMe in combination with one ormore “absorption enhancers”. “Absorption enhancers,” such asparacellular permeability enhancers (PPE) or “promoters” typically fallinto the broad chemical categories of detergents or surfactants (such aszwitterionic surfactants), non-surfactants (such as unsaturated cyclicureas), fatty acids, bile acids and chelating agents. Each agent mayimprove absorption of orally delivered active ingredients, by one ormore mechanisms exemplified by altering the rheology of the overlyingmucous, fluidizing the cell membrane lipid bilayer, affecting the tightjunctional complex, inhibiting enzyme or transporter activity,influencing the drug itself in some way, among others. In certainembodiments, absorption enhancers used herein may function through anumber of chemical or physical interactions including those that: (1)modulate SAMe solubility; (2) improve SAMe mucous diffusivity; (3)protect SAMe from pH, lumenal and/or brush border enzymes; (4) protectSAMe from nonspecific binding sites; and (5) improve SAMe's permeabilitythrough the mouth and/or gastrointestinal epithelium.

Examples of absorption enhancers which are suitable for use in thepresent embodiments include, but are not limited to, small moleculeenhancers that are commonly referred to as CPEs (chemical penetrationenhancers; as listed in Table 1 below), bile salts, surfactants,phospholipids, glycerides and fatty acids, as well as peptide hormones,cytoskeletal perturbing agents, oxidants, calcium ion (Ca⁺⁺) chelatorsand ionophores.

TABLE 1 List of CPEs CAS Abbreviations Chemical Name Category number SLSSodium lauryl sulfate AS 151-21-3 SDS Sodium decyl sulfate AS 142-87-0SOS Sodium octyl sulfate AS 142-31-4 SLA Sodium laureth sulfate AS68585-34-2 NLS N-Lauryl sarcosinate AS 137-16-6 CTAB Cetyltrimethylammonium bromide CS 57-09-0 DTAB Decyltrimethyl ammonium bromide CS2082-84-0 BDAC Benzyldimethyl dodecyl ammonium chloride CS 139-07-1 TTACMyristyltrimethyl ammonium chloride CS 4574-04-3 DPC Dodecyl pyridiniumchloride CS 104-74-05 ASB Amidosulfobetaine-16 or 3-[N,N-Dimethyl(3- ZS52562-29-5 palmitoylaminopropyl) ammonio]-propanesulfonate DPSDecyldimethyl ammonio propane sulfonate ZS 15163-36-7 CHAPS3-[(3-Cholamidopropyl)dimethylammonio]-1- ZS 75621-03-3 propanesulfonateCHAPSO 3-[(3-Cholamidopropyl)dimethylammonio]-2- ZS 82473-24-3hydroxy-1-propanesulfonate ADPS Alkyl dimethylammoniopropane sulfonate(general ZS N/A class) SB-163-(N,N-Dimethylpalmitylammonio)propanesulfonate ZS 2281-11-0 MPSMyristyldimethyl ammonio propane sulfonate ZS 14933-03-6 PPSPalmityldimethyl ammonio propane sulfonate ZS 2281-11-0 AB Amidobetaine(general class) ZS N/A CAPB Cocamidopropyl betaine or {[3- ZS 86438-79-1(Dodecanoylamino)propyl](dimethyl)ammonio}acetate Betaine ZS 107-43-7DDN N,N-Dimethyl-N-dodecylglycine betaine ZS 89045-97-6 CBC ChemBetaineCAS ZS N/A mixture CAHS Cocamidopropyl hydroxysultaine ZS 68139-30-0 CBOChemBetaine Oleyl ZS N/A mixture AO Amine Oxide (general class) ZS N/ALDAO N,N-dimethyldodecylamine N-oxide ZS 1643-20-5 PCC Palmitoylcarnitine chloride ZS 6865-14-1 IP Nonylphenoxypolyoxyethylene NS68412-54-4 T20 Polyoxyethylene sorbitan monolaurate NS 9005-64-5 T40Polyoxyethylene sorbitan monopalmitate NS 9005-66-7 SP80 Sorbitanmonooleate NS 1338-43-8 TX100 Triton-X-100 NS 9002-93-1 SDC Sodiumdeoxycholate BS 302-95-4 SGC Sodium glycocholate BS 863-57-0 CA CholicAcid BS 81-25-4 HA Hexanoic Acid FA 142-62-1 HPA Heptanoic Acid FA111-14-8 LME Methyl Laurate FE 111-82-0 IPM Isopropyl myristate FE110-27-0 IPP Isopropyl palmitate FE 142-91-6 MPT Methyl palmitate FE112-39-0 SDE Dibutyl sebacate FE 109-43-3 SOA Sodium oleate SS 143-19-1UR Urea FM 57-13-6 LAM Lauryl amine FM 124-22-1 CL Caprolactam NR105-60-2 MP Methyl pyrrolidone NR 872-50-4 OP Octyl pyrrolidone NR2687-94-7 MPZ Methyl piperazine NR 109-01-3 PPZ Phenyl piperazine NR92-54-6 EDTA Ethylenediaminetetraacetic acid OT 60-00-4 SS Sodiumsalicylate OT 54-21-7 CP Carbopol 934P OT 9003-04-7 GA Glycyrrhetinicacid OT 471-53-4 BL Bromelain OT 9001-00-7 PO Pinene oxide OT 1686-14-2LM Limonene OT 5989-27-5 CN Cineole OT 470-82-6 ODD Octyl dodecanol OT5333-42-6 FCH Fenchone OT 7787-20-4 MTH Menthone OT 14073-97-3 TPMBTrimethoxy propylene methyl benzene OT 2883-98-9 AS Anionic surfactants,CS cationic surfactants, ZS zwitterionic surfactants, NS nonionicsurfactants, BS bile salts, FA fatty acids, FE fatty esters, FM fattyamines, SS sodium salts of fatty acids, NR nitrogen-containing rings, OTothers.

Recent advances in drug absorption research led to the discovery of anincreasing number of integral membrane, adaptor, regulator and signalingproteins in tight and adherens junctions. Tight junctions areintercellular junctions between cells that form a barrier between thecells. In this way, materials (e.g. small molecules, proteins and drugs)cannot pass between cells but rather must be taken up by the cell andthus enables the cells to regulate what is allowed through. Tightjunctions occur in many regions throughout the body including the mouth,small intestine, large intestine and colon and vary in density/tightnesswithin different regions. Within the GI tract, tight junctions refer tothe areas between adjacent endothelial cells and act, in part, toregulate the uptake of digested materials. Tight junctions are highlyregulated and are one of the key elements that form the barrier betweenthe luminal environment of the mouth and/or GI tract and the rest of thebody.

Tight junctions have three main functions: (1) to hold cells together,(2) to block the movement of integral membrane proteins between theapical and basolateral surfaces of the cell, allowing the specializedfunctions of each surface (for example receptor-mediated endocytosis atthe apical surface and exocytosis at the basolateral surface) to bepreserved (this aims to preserve the transcellular transport) and (3) toprevent the passage of molecules and ions through the space betweencells and therefore materials must actually enter the cells (bydiffusion or active transport) in order to pass through the tissue. Thispathway provides control over what substances are allowed through.

New tight junction modulators or opening agents are currently underdevelopment, which can directly target tight or adherens junctionproteins, the signaling pathways regulating junctional function, ortight junction associated lipid raft microdomains. Modulators actingdirectly on tight junctions include peptides derived from zonulaoccludens toxin, Clostridium perfringens enterotoxin, peptides selectedby phage display that bind to integral membrane tight junction proteins,and lipid modulators. They can reversibly increase paracellulartransport and drug delivery and have a potential to be used aspharmaceutical excipients to improve drug delivery across epithelialbarriers and the blood-brain barrier. Exemplary “tight junctionmodulators” suitable for use in the present embodiments include, but arenot limited to, chitosan, poly(acrylic acid), cytochalasin D; caprate,spermine, taurocholate (including sodium and other salt forms) and otherbile acids and/or their salts (such as cholic acid, sodium cholate orpotassium cholate), as well as more recently identified agents whichinclude peptides derived from zonula occludens toxin or Clostridiumperfringens enterotoxin. Classes of tight junction modulators includedherein thus include: saturated and/or unsaturated fatty acids or theircorresponding carboxylate salts (e.g. C6-C24 fatty acids, or carboxylatesalts thereof, especially C8-C22 fatty acids, or carboxylate saltsthereof, C10-C20 fatty acids or carboxylate salts thereof, C6-, C7-,C8-, C9-, C10-, C11-, C12-, C13-, C14-, C15-, C16-, C17-, C18-, C19-,C20-, C21-, C22-fatty acids or carboxylate salts thereof), saturated andunsaturated sulfonic acids and sulfonate salts thereof (e.g. C6-C24sulfonic acids or sulfonate salts, especially C8-C22 sulfonic acids orsulfonate salts, C10-C20 sulfonic acids or sulfonate salts, C8-, C9-,C10-, C11-, C12-, C13-, C14-, C15-, C16-, C17-, C18-, C19-, C20-, C21-,C22-sulfonic acids or sulfonate salts); zwitterionic surfactants (e.g.3-(N,N-Dimethylpalmitylammonio)propanesulfonate, decyldimethyl ammoniopropane sulfonate, myistyldimethyl ammonio propoane sulfonate,cocamidopropyl hydroxysultaine (ChemBetaine® CAS), oleyl betaine(ChemBetaine®Oleyl), or palmitoyl carnitine chloride); fatty amines(e.g. C6-C24 fatty amines, especially C8-C22 fatty amines, C10-C20 fattyamines, C6-, C7-, C8-, C9-, C10-, C11-, C12-, C13-, C14-, C15-, C16-,C17-, C18-, C19-, C20-, C21-, C22-fatty amines), as well as otherorganic acids (e.g. tartaric acid) and cyclodextrins (e.g.alpha-cyclodextrin, beta-cyclodextrin, or gamma-cyclodextrin). Exemplaryfatty acids that may be used include hexanoic, heptanoic, capric, lauricacid, myristic acid, palmitic acid, stearic acid, arachidic acid,myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, linoleicacid, α-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucicacid, docosahexaenoic acid. Exemplary carboxylate salts that may be usedinclude sodium or potassium caprate, caprylate, laurate, myristate,palmitate, stearate, arachidate, myristoleate, palmitoleate, sapienate,oleate, linoleate, α-linolenate, arachidonate, eicosapentaenoate,erucate, docosahexaenoate. Specific carboxylate salts include sodiumcaprate, sodium caprylate, and sodium laurate. Specific fatty aminesthat may be used include lauryl amine (N-dodecylamine), decylamine,nonylamine, octylamine, heptylamine or hexylamine. Exemplary sulfonicacids that may be used include octane sulfonic acid, decane sulfonicacid (e.g. sodium 1-decanesulfonate), dodecane sulfonic acid,tetradecane sulfonic acid, hexadecane sulfonic acid, octadecane sulfonicacid, eicosane sulfonic acid, docosane sulfonic acid or tetracosanesulfonic acid. Specific sulfonic acids that may be mentioned includedioctyl sodium sulfosuccinate.

Zwitterionic agents, for example zwitterionic surfactants as listedabove, are preferred absorption enhancing agents in some embodiments.These agents are compounds with a net neutral charge, which have atleast one positive and one negative electrical charge on different atomswithin the molecule. They are sometimes referred to as inner salts ordipolar ions. Zwitterionic agents for use in certain embodiments aregenerally neutral at physiologically relevant pH levels (i.e. about pH6-8). A preferred zwitterionic surfactant for use in the certainembodiments is an acyl carnitine, more preferably, a long chain acylcarnitine, and even more preferably, palmitoyl carnitine. Some of theseagents are naturally occurring molecules that are intermediatestructures, formed by the body to assist in the intracellular transportof long chain acyl groups to the mitochondria for energy production,though synthetic analogs may also be prepared. See the formula below forthe general structure as well as other examples of long chain acylcarnitines which may be used in the present embodiments:

Name R Structure R = Stearoyl carnitine

C₁₇H₃₅ Palmitoyl carnitine

C₁₅H₃₁ Myristoyl carnitine

C₁₃H₂₇ Lauroyl carnitine

C₁₁H₂₃ Decanoyl carnitine

C₉H₁₉

Thus, some embodiments comprise a combination of SAMe and one or moreacyl carnitines of the above formula in which the R is CH₃(CH₂)_(n), nis 7-21, preferably 9-17, and particularly 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20 or 21.

Certain embodiments thus relate to examining the effect of acylcarnitine chain length in in vitro cell permeability of SAMe inappropriate models (e.g. Caco-2) and also the effect of chain length onthe pharmacokinetic properties in animal models (e.g. Beagle dogs) insolid or semi-solid oral dosage forms.

Another example of zwitterionic surfactants included for use in thepresent embodiments are the long alkyl chain N substituteddimethylammoniopropane sulfonates (Nsubstituted-N,N-dimethyl-3-ammonio-1-propane sulfonates). Thesemolecules are sometimes referred to as the sulfobetaines. See theformula below for the general structure as well as other examples oflong chain dimethylammoniopropane sulfonates which may be used in thepresent embodiments:

Name R Structure R = Sulfobetaine 18

C

H

Sulfobetaine 16

C

H

Sulfobetaine 14

C

H

Sulfobetaine 12

C

H

Sulfobetaine 10

C

H

indicates data missing or illegible when filed

Thus, some exemplary embodiments comprise a combination of SAMe and oneor more dimethylammoniopropane sulfonates of the above formula in whichthe R is CH₃(CH₂)_(n), n is 7-21, preferably 9-19, and particularly 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21.

Other examples of zwitterionic surfactants included for use in thepresent embodiments are listed with their general structures in thetable below:

zwitterionic/amphoteric surfactant General Structure alkyl betaines

Alkyl lamidobetaines

Alkyl amine oxides

3-[N,N-Dimethyl(3-acylaminopropyl) ammonio]-propanesulfonates

alkylamidopropylhydroxysultaines

Thus, some embodiments comprise a combination of SAMe and one or more ofthese zwitterionic surfactants of the formulas shown in the table above,in which the R is CH₃(CH₂)_(n), n is 7-21, preferably 9-19, andparticularly 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21.

The inventors show for the first time that a subset of high potencytight junction opening absorption enhancers, which increase SAMeabsorption when used at very low levels, exists. Furthermore,identification of potential high potency tight junction opening agentsmay be carried out using, for example, a SAMe Caco-2 cell model asdescribed in the Examples. Based on the concentration of tight junctionmodulator utilized as well as the increase in SAMe permeabilityachieved, suitable high potency tight junction modulators elicit atleast a two-fold increase in SAMe uptake in vitro when used at lowconcentrations (i.e. about 2 mM or below). More preferably, at least athree-fold increase in SAMe uptake occurs at low concentrations of thesetight junction opening agents and, even more preferably, a four-fold orgreater increase is achieved.

Other zwitterionic surfactants for use in the present embodiments alsohave long alkyl chains as a structural feature and particular groups ofmolecular analogs, differing only in alkyl chain length, will beinvestigated.

Thus certain exemplary embodiments relate to compositions comprising aphysiologically effective dosage of SAMe and at least one zwitterionicsurfactant. In some exemplary embodiments said zwitterionic surfactantis an acyl carnitine. In other exemplary embodiments said zwitterionicsurfactant is an N substituted dimethylammoniopropane sulfonate orsulfobetaine. In preferred embodiments, said zwitterionic surfactant isco-formulated with the physiologically acceptable dosage of SAMe. Otherexemplified embodiments relate to compositions comprising aphysiologically effective dosage of SAMe and at least one high potencytight junction opening enhancer.

In some embodiments, the solid oral composition contains aphysiologically effective dose of S-adenosylmethionine (SAMe) and atight junction opening enhancer, which enhances delivery of SAMe, asmeasured by an increase in SAMe Cmax and/or AUC. The increase in SAMeCmax may be determined by comparison of SAMe delivery of the compositioncomprising SAMe and the absorption enhancer (test dosage) with deliveryof a SAMe using a comparable control, which contains the same amount ofSAMe but lacks the absorption enhancer. In some embodiments, thecomposition comprising SAMe and the absorption enhancer is substantiallysimilar to the control, with the exception that the control lacks theabsorption enhancer. (In this context, “substantially similar” meansthat the test dosage and the control contain the same amount of SAMe inthe same dosage form with the same types of excipients.) Likewise, theincrease in AUC may be determined by comparing SAMe delivery using thecomposition comprising SAMe and the absorption enhancer with delivery ofa SAMe using the control. In some embodiments, the SAMe Cmax obtainedwith the composition comprising SAMe and enhancer may be at least 110%,at least 120%, at least 130%, at least 140%, at least 150%, at least160%, at least 180%, at least 200%, at least 300%, at least 400% or atleast 500% of the SAMe Cmax obtained with the control. Likewise, in someembodiments the SAMe AUC obtained with the composition comprising SAMeand enhancer may be at least 110%, at least 120%, at least 130%, atleast 140%, at least 150%, at least 160%, at least 180%, at least 200%,at least 300%, at least 400% or at least 500% of the SAMe AUC obtainedwith the control. In certain embodiments, the SAMe Cmax and/or AUC inthe presence of a tight junction opening enhancer is at least 10%, atleast 20%, at least 30%, at least 40%, at least 50%, at least 60%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, at least 100%, at least 150%, at least 200%, at least 300%,at least 400%, or at least 500% higher than the Cmax of the controlgroup. In certain embodiments, the SAMe Cmax and/or AUC in the presenceof a tight junction opening enhancer is at least 2-fold, at least3-fold, at least 4-fold, at least 5-fold, at least 10-fold higher thanthe Cmax of the control group. In some embodiments, the Cmax and/or AUCare determined in a suitable test population comprising at least sixsubjects without outliers. In some embodiments, the suitable testpopulation is a population of mammals, such as humans, dogs, cats, rats,mice, monkeys or other mammalian test subjects. In some embodiments, thetest subjects are humans. In some embodiments, the test subjects aredogs.

Pharmacokinetic parameters such as average maximum plasma concentrationof SAMe (Cmax) are determined using a bioanalytical method with adequatesensitivity, specificity, ruggedness, stability and repeatability (forexample, a qualified liquid chromatography triple quad mass spectrometrybased method coupled with a suitable extraction method for theseparation of analyte from plasma). SAMe AUC values were calculated from0-24 hours using the trapezoid method and are uncorrected for baseline,endogenous SAMe levels. A “suitable test population” preferably has sixor more subjects who are dosed fasted. All members of the “suitable testpopulation” have pharmacokinetic parameters for SAMe that fall withinstatistically normal ranges (i.e. no outliers) and no member will beincluded on the basis of non-standard or unusual SAMe absorption ormetabolism. The average Cmax values are preferably derived by averagingthe individual Cmax concentrations for each member of the subject group.Use of present compositions and methods in vivo provides high SAMe Cmaxand/or AUC values in comparison to conventional dosage forms of SAMe.For greater clarity all references to dose within this patent refer todose as the dose of SAMe ion.

Tight junction modulators may be particularly effective in improvingmodified release dosage forms targeting more distal segments of the GItract. The porosity of tight junctions is tighter in distal segmentssuch as the ileum and colon, compared to, for example, the duodenum(i.e. tight junctions of the duodenum are more porous than those of thelower GI segments.) In addition, transit time in the upper GI tract isfaster than in the lower GI tract. The combinations of less porous tightjunctions coupled with the slower transit time in the lower segmentssuggest that the use of tight junction modulators in the lower GI tractmay be more impactful on a relative basis. This effect could be expectedto extend SAMe delivery (as well as increase its absorption using tightjunction modulators) in the colon and in the case of suppositoryformulation, to the rectum, which would constitute targeted delivery toa desired site of action in the treatment of colon and/or other gastriccancers. This distally targeted delivery is optimal to exploit theantineoplastic properties of SAMe on colon and/or other gastric cancercells. Thus in some embodiments, compositions comprising a delayedrelease component, such that the release of SAMe is configured todissolve in the large intestine or colon, are included. Otherembodiments include methods of treating a condition related to the colonor large intestine, which condition is amenable to treatment with SAMe,comprising administering to a patient a physiologically effective amountof a SAMe and at least one absorption enhancing agent. Preferably, saidabsorption enhancing agent is a tight junction modulating agent.

Improved buccal delivery of SAMe is also considered practical usingformulations provided herein comprising one or more tight junctionmodulators considering the presence of tight junctions in the mouth.Thus non-parenteral formulations provided herein are meant to includethose which target buccal, upper and lower intestinal regions includingthe colon and rectum.

Thus in certain embodiments, provided herein are compositions for buccaldelivery comprising a physiologically acceptable dosage of SAMe and atleast one tight junction modulator.

In some embodiments the composition is administered as a buccal dosageform. In other embodiments the composition is administered as asuppository.

In certain embodiments, provided herein is a lower GI targetedmodulator-enhanced component in combination with an upper GI-targetedconventional or modulator-enhanced component to create a modifiedrelease dosage form with enhanced absorption over an extended period oftime.

Preferably, the suitability of a particular “absorption enhancer”,including “tight junction modulators,” will be identified in vitro byuse of SAMe cellular permeability studies. Most relevant cell lines willsuffice for such in vitro experimentation including, but not limited to,Caco-2 cells (as described in Example 3). In addition, the use ofreferences in the art may also provide insight into potentially suitable“absorption enhancers” or “tight junction modulators” for use in thepresent embodiments.

Nanocarriers to Increase Delivery of SAMe

In certain embodiments, encapsulating SAMe into nano-sized carrierswhich are suitable for use in non-parenteral administration of SAMe(e.g. nanoparticles and colloidal systems) may result in increaseddelivery to the cells. Several approaches have been described thatappear to increase transcellular intestinal absorption without damagingthe epithelium. These approaches can be categorized into methods thatstabilize the drug, increase drug solubility or alter itscharacteristics to improve transcellular permeability. Various colloidalsystems which may be suitable for enhancing the absorption of SAMe areexemplified by sub-micron emulsions, polymeric nanoparticles,microparticles, and the like. There are various physicochemical factorsgoverning gastrointestinal uptake of such systems including size, sizedistribution, consistency, hydrophobicity and surface properties whichmay be modulated in order to enhance SAMe cellular uptake.

Dosing with Formulations Exhibiting Enhanced-Absorption andBioavailability of SAMe

In some embodiments the enhanced-absorption SAMe formulations providedherein relate to enhanced nutritional support, or dietary supplementhealth improvements including, but not limited to, mood improvement,joint health and liver function. In some exemplary embodiments thedisorder is related to the dietary management of a disease throughadditional supplementation of SAMe which cannot be reached through diet(e.g. a “medical food”.)

Some exemplary embodiments relate to a method for treating and/orprophylaxis in a subject a disease or disorder selected from the groupconsisting of, but not limited to, a mental or psychiatric disorder(e.g. psychotic/mood or non-psychotic mental disorders exemplified bydepression and substance related disorders, respectively), a nervoussystem disease/disorder (e.g. a central nervous system diseaseexemplified by Alzheimer's, Progressive Supranuclear Palsy or otherTauopathy disorders), other neurological disease/disorders (e.g.headaches and sleep disorders), conditions associated with injury to thecentral nervous system, a liver disease/disorder (e.g. alcoholic liverdisease), a cancer (e.g. solid and blood-borne cancers), a jointdisease/disorder (e.g. arthritis), an inflammatory disease/disorder(e.g. ulcerative colitis), an autoimmune disease/disorder (e.g. systemiclupus erythematosis and rheumatoid arthritis), a degenerativedisease/disorder (e.g. Amyotrophic Lateral Sclerosis), a soft-tissuedisease/disorder (e.g. a fibromyalgia disorder), a paindisease/disorder, a genetic disorder related to hyper- orhypo-methylation, a gastrointestinal disease/disorder, a cardiovasculardisease/disorder, and a disorder induced in whole or in part byoxidative or free-radical damage, comprising administering to saidsubject an exemplary composition which enhances the absorption andbioavailability of a physiologically effective amount of exogenous SAMe.

Some embodiments relate to therapeutic use of the exemplary compositionsdisclosed herein for treatment of a mental or psychiatric disorderselected from the group consisting of anxiety disorders, depressivedisorders, eating disorders, bipolar disorder, abuse disorders,dependence disorders, Axis II disorders, and psychosis. In someexemplary embodiments, the mental or psychiatric disorder is an anxietydisorder selected from the group consisting of generalized anxietydisorder, posttraumatic stress disorder, social anxiety disorder, panicdisorder, Schizophrenia and obsessive compulsive disorder. In someexemplary embodiments, the mental or psychiatric disorder is adepressive disorder selected from the group consisting of majordepressive disorder, multi-infarct dementia, minor depression,postpartum or late-life depression (and the like), Parkinson'sdepression, HIV-associated depression, brief recurrent depression,dysthymia or depression NOS (Not Otherwise Specified). In some exemplaryembodiments, the mental or psychiatric disorder is an eating disorderselected from the group consisting of bulimia nervosa, anorexia nervosa,binge eating disorder, obesity, or eating disorder NOS. In someexemplary embodiments, the mental or psychiatric disorder is bipolardisorder, an abuse disorder or a dependence disorder, including abuseof, or dependence on, alcohol, nicotine, cocaine, codeine, oxycodone,hydrocodone or other opiates. In some exemplary embodiments, the mentalor psychiatric disorder is an Axis II disorder selected from borderlinepersonality disorder.

In some exemplary embodiments, the disorder is a nervous systemdisorder, including a central nervous system (CNS) disorder such asParkinson's disease, Alzheimer's disease, Tauopathy disorders (such as,for example, Progressive Supranuclear Palsy (PSP)), Angelman Syndrome(genetic disorder), Multiple Sclerosis (MS) and pre-dementia and/orcognitive impairment.

In some exemplary embodiments, the disorder is a comorbid disorder, suchas comorbid depression arising in a subject who is undergoing treatmentfor one or more diseases or disorders such as but not limited to,cancer, Parkinson's, cardiovascular disease and HIV. In certainembodiments the comorbid disorder is caused by one or more therapiesbeing utilized to treat said one or more diseases or disorders.

In some exemplary embodiments, the disorder is a drug-induced disorderor side-effect, such as anti-depressant-induced sexual dysfunction orinsomnia.

In some exemplary embodiments, the disorder is a result of an injury tothe CNS such as spinal cord injury or brain damage, memory loss,cognitive impairment and/or learning disability.

In some exemplary embodiments, the disorder is a liver disorder selectedfrom the group consisting of alcoholic liver disease, fatty liverdisease (non-alcoholic) hepatitis (both viral and non-viral), livercancer, oxidative liver disease, HISS-dependent insulin resistance,cholestasis and cirrhosis.

In some exemplary embodiments, the disorder is a cancer selected fromthe group consisting of cancers occurring in one or more of the liver,colon, rectum, ovaries, urethra, testicles, bladder, breast, stomach,esophagus, pancreas, head and neck, lung, blood, skin (such as actinickeratosis, basal cell cancer, superficial basal cell cancer, squamouscell cancer, and melanoma) and adenocarcinomas.

In some exemplary embodiments, the disorder is a joint disorder such as,for example, arthritis and osteoarthritis.

In some exemplary embodiments, the disorder is an inflammatory disorderselected from the group comprising systemic lupus erythematosis, Reye'ssyndrome, rheumatic fever, allergic rhinitis, myasthenia gravis,temporal arteritis, vasculitis, psoriasis, atopic dermatitis, rosacea,eczema, alopecia universalis, scleroderma, pemphigus, contactdermatitis, ankylosing spondylitis, dermatomyositis, polymyositis,celiac sprue, Guillain-Barré syndrome, multi-infarct dementia,post-cerebral vascular accident reperfusion damage, Addison's disease,Hashimoto's thyroiditis, asthma, upper respiratory inflammationsymptoms, chronic bronchitis, atherosclerosis, pernicious anemia,autoimmune hepatitis, prostatitis, pelvic inflammatory disease,Goodpasture's syndrome, Wegener's granulomatosis, chronic nephritis,Sjogrens syndrome, or allergic conjunctivitis.

In some exemplary embodiments, the disorder is a gastrointestinaldisorder such as inflammatory bowel disease (IBD), Crohn's disease orulcerative colitis (UC).

In some exemplary embodiments, the disorder is a soft tissue diseasesuch as fibromyalgia.

In some exemplary embodiments, the disorder is a pain disorder such asfibromyalgia, chronic headaches, shingles, reflex sympathetic dystrophyand polyneuropathy.

In some exemplary embodiments, the disorder is a cardiovascular disorderwhich is related to hyper- or hypo-homocysteinemia such as coronaryheart disease, stroke, peripheral vascular disease and atheroscleroticdisease.

In some exemplary embodiments, the disorder is related to a genetic ormedical condition related to a deficiency of the methylation pathwaysuch as methylenetetrahydrofolate reductase deficiency.

In some exemplary embodiments, the disorder is a metabolic disorder,such as type 2 diabetes.

In some exemplary embodiments, the etiology of the disorder may includeoxidative or free-radical damage, and is selected from the groupcomprising chronic fatigue syndrome, temporal arteritis, vasculitis,multi-infarct dementia, chronic emphysema, or chronic nephritis.

Among the advantages provided by enhanced-absorption SAMe formulationsof the present embodiments, included are the convenience and concomitantimproved subject compliance due to reduced daily dosing, an improvedside-effect profile (such as decreased stomach irritation andpotentially decreased tendency to induce mania in manic depressivesubjects or subjects at risk for manic episodes) and other side effectsassociate with or caused by the relatively high doses of SAMe (typicallyabout 400 to about 3200 mg SAMe ion/day, more typically about 800 toabout 1600 mg SAMe ion/day) necessary to achieve a desired effect.

As used herein, the term “desired effect” includes a “therapeuticeffect”, “pharmaceutical effect”, “dietary effect” (e.g. for use as amedical food), “nutraceutical effect” and “nutritional effect”. Thus, a“desired effect” includes ameliorating at least one symptom of aphysiological disorder or disease state in a subject, or improving atleast one performance variable (such as improved concentration, memory,mood, nutrition status or liver status) when used as a nutritionalsupplement in a subject. The “desired effect” may be achieved throughnutritional supplementation using SAMe formulations provided herein orthrough administration using a clinical, pharmaceutical or veterinarydosing regimen of SAMe formulations provided herein.

Suitable subjects for dosing according to the methods and compositionsprovided herein include warm-blooded mammals such as humans, domestic orexotic animals or livestock; domesticated avian subjects such aschickens and ducks; and laboratory animals suitable for research use.When used for treating a disease or disorder in a subject, varioussymptoms of specific physiological disorders and disease states arecontemplated as being treatable within the context of the presentembodiments and details of which are set forth below. However, it is tobe recognized that the understanding of various disease states by thoseof skill in the art is not static and this is the same for performancevariables related to nutritional supplementation. Thus, though thedescription above is intended to be illustrative of the variousdisorders, disease states, symptoms or performance variables that may betreated using the enhanced-absorption SAMe formulations according to thepresent embodiments, a person skilled in these arts will be expected toapply such knowledge.

Dosing with Multiple Dosing Units

Some exemplary embodiments provided herein relate to treatment of and/orprophylaxis of one or more diseases in a subject, wherein the treatmentof and/or prophylaxis of one or more diseases and/or disorders comprisesadministering to the subject an absorption-enhanced formulationcomprising a physiologically acceptable dosage of S-adenosyl methionine(SAMe), or a proprietary salt thereof.

Some other exemplary embodiments provided herein relate to SAMenutritional supplements and/or dietary supplements for improvement ofone or more nutritional performance variables in a subject, wherein thenutritional performance variables are one or more of concentration,memory, mood, nutritional status and liver status, and wherein anabsorption enhanced formulation comprising a physiologically acceptabledosage of 5-adenosyl methionine (SAMe), or other proprietary SAMe saltsthereof, is administered to a subject.

In some exemplary embodiments, the absorption-enhanced SAMe may bedivided between multiple daily doses. Multiple daily doses need not beidentical and may comprise one or more dosage forms in combination. Insome exemplary embodiments, the enhanced-absorption SAMe may be dividedinto two or more daily doses. Each dose may be administered as a singledosage unit exemplified by, a single tablet, capsule or caplet, oralternatively may be divided into multiple dosage units. In someembodiments, a twice-daily dose of from about 100 to about 1600 mg ofSAMe ion per dose may be divided into one to four dosage units of fromabout 100 to about 800 mg of SAMe ion per unit. In each case, the formof the dosage unit may be a capsule, a tablet, a caplet (single ormulti-compartment) or an extended release dosage unit and the like. Insome embodiments, the absorption enhancer and SAMe are provided in aoral dosage form wherein separate compartments of the oral dosage formcontain either the absorption-enhancing agent or SAMe. In otherembodiments the absorption-enhancing technology is administeredseparately from the SAMe dosage form. Preferably, the oral dosage formis a tablet, capsule or gel-capsule.

In some embodiments, the ratio (weight:weight) of S-adenosylmethionineto tight junction opening absorption enhancer is from 1:100,000 to100,000:1. In other embodiments, the ratio of S-adenosylmethionine totight junction opening absorption enhancer is from 1:150 to 150:1, from1:10 to 10:1, from 1:5 to 10:1, from 1:2 to 10:1, from 1:1 to 10:1, from1:5 to 5:1, or from 1:2 to 2:1. In some embodiments, the ratio ofS-adenosylmethionine to tight junction opening absorption enhancer is1:1.

Conventional SAMe dosing generally administers up to 1600 mg of SAMe ionper day bi-daily (BID) in order to achieve maximum activity of the drug.Tablets are most often available commercially in 200 mg and 400 mg dosesSAMe ion which require subjects to ingest 4-8 tablets per day. This isinconvenient with respect to the amount of time needed as well as thepotential error in consistent dosing (i.e. if a dose is missed).Provided herein are novel compositions and methods which reduce theeffective dose of SAMe (i.e. reduce the number of tablets necessary in aday to achieve the same or better efficacy as compared to conventionaldosing regimens) and/or eliminate the need to dose bi-daily. Byimproving SAMe absorption, a new method of SAMe therapy is availablewhich lowers the amount of SAMe dose required to elicit an effectiveresponse by providing compositions comprising one or moreabsorption-enhancing technologies. These exemplary “low dose”formulations may provide a lower daily pill count which is beneficial tothose taking SAMe as it will reduce the time, cost and inconvenience ofself-administering large doses.

Some exemplary embodiments relate to administration of the selectedphysiologically acceptable dosage on a once-a-day basis. In someembodiments, the once-a-day dose may be administered in a single dosageunit exemplified by, a single tablet, capsule, or caplet. In otherexemplary embodiments, the single dose may be administered as multipletablets, capsules or caplets taken at one time. In some embodiments, forinstance, a dosage of about 400 to 3200 mg of SAMe per day may bedivided into two, three, four or more tablets, capsules or caplets ofabout 50 to 2000, preferably about 100 to 1600 mg of SAMe per unit. Insome preferred embodiments, the daily dose may comprise two, three orfour units (e.g. tablets, capsules or caplets) of about 100 to 800 mg ofSAMe ion per unit. Suitable dosage regimens included are: four units ofabout 50-400 mg of SAMe ion per unit, e.g. 50, 100, 150, 200, 250, 300,350 or 400 mg SAMe ion per unit; three units of about 50-1000 mg of SAMeion per unit, e.g. 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550,600, 650, 700, 750, 800, 850, 900, 950 or 1,000 mg of SAMe ion per unit;two units of about 50-1600 mg of SAMe ion per unit, e.g. about 50, 100,150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800,850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400,1450, 1500, 1550 or 1600 mg of SAMe ion per unit.

Some exemplary embodiments provided herein relate to “low-dose” SAMecompositions. By increasing the bioavailability of exogenous SAMe, thedaily administered dose of SAMe may be substantially lowered byadministration of compositions with improved SAMe absorption. Theseexemplary “low-dose” treatments may enable a lower daily pill count.

Fed vs. Fasting Dose

In some embodiments, it may be advantageous to ensure that the subjectis either fed or fasted (e.g. overnight for at least about 6, especiallyabout 8, hours). It is considered that food administered at the sametime, immediately (i.e. less than about 30, especially less than about15 minutes) before or soon (e.g. less than about 10 minutes) after theabsorption enhanced SAMe formulation provided herein is administered tothe subject may increase or decrease the rate of gastric emptying andthus affect the rate of uptake of SAMe from the formulation. Thus, insome embodiments, provided herein are methods of administering theabsorption enhanced SAMe formulation with food, wherein food is ingestedeither before or during SAMe treatment.

Combinations of SAMe with Other Active Ingredients

Some exemplary embodiments provided herein relate to combinations ofSAMe with one or more active ingredients that can be prescribed or usedfor treating and/or prophylaxis in a subject a disease or disorderselected from the group consisting of, but not limited to, a mental orpsychiatric disorder (e.g. psychotic or non-psychotic mental disorderssuch as depression and substance abuse disorders, respectively), anervous system disease/disorder (e.g. a central nervous system diseasesuch as Alzheimer's, Progressive Supranuclear Palsy, or other Tauopathydisorders), other neurological disease/disorders (e.g. headaches andsleep disorders), conditions associated with injury to the centralnervous system, a liver disease/disorder (e.g. alcoholic liver disease),a cancer (e.g. solid and blood-borne cancers), a joint disease/disorder(e.g. arthritis), an inflammatory disease/disorder (e.g. ulcerativecolitis), an autoimmune disease/disorder (e.g. systemic lupuserythematosis and rheumatoid arthritis), a degenerative disease/disorder(e.g. Amyotrophic Lateral Sclerosis), a soft-tissue disease/disorder(e.g. a fibromyalgia disorder), a pain disease/disorder, a geneticdisorder related to hyper or hypo methylation, a gastrointestinaldisease/disorder, a cardiovascular disease/disorder, and a disorderinduced in whole or in part by oxidative or free-radical damage,comprising administering to said subject an exemplary compositionprovided herein which enhances the absorption and bioavailability of aphysiologically effective amount of exogenous SAMe.

In some exemplary embodiments provided herein relate to combinations ofSAMe with one or more active ingredients that can be prescribed or usedfor treatment of and/or prophylaxis of mental or psychiatric disordersin a subject include, but are not limited to, tricyclic antidepressants(TCAs), tetracyclic antidepressants, aminoketones, phenylpiperazines,selective serotonin reuptake inhibitors (SSRIs), monoamine oxidaseinhibitors (MAOIs), serotonin-norepinephrine reuptake inhibitors(SNRIs), norepinephrine-serotonin reuptake inhibitors (NSRIs), dopaminereuptake inhibitors, norepinephrine-dopamine reuptake inhibitors,norepinephrine reuptake inhibitors, selective serotonin reuptakeenhancers, noradrenergic and serotonin specific antidepressants,substance P receptor antagonists, neurokinin receptor antagonists suchas saredutant, corticotrophin release factor antagonists such asmifepristone, atypical antipsychotics such as aripiparazole, commonlyused antidepressant augmenters such as lithium or triple reuptakeinhibitors.

Some exemplary embodiments provided herein relate to combinations ofSAMe with one or more device therapies that can be prescribed or usedfor treatment of and/or prophylaxis of mental or psychiatric disordersin a subject include, but not limited to ECT (electro convulsivetherapy) and electric shock therapy.

Some exemplary embodiments provided herein relate to combinations ofSAMe with one or more active ingredients that can be prescribed or usedfor treatment of and/or prophylaxis of a nervous system disease/disorderin a subject include, but are not limited to anticonvulsants such aspregabalin, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)receptor antagonists, methylphosphonate (NMPA) receptor antagonists,histamine receptor antagonists, nitric oxide (NO) modulators, glutamatereceptor antagonists, acetylcholinesterase inhibitors, dopamineagonists, N-methyl-d-aspartate (NMDA) receptor antagonists such asmemantine, cholinesterase inhibitors such as donepezil,neuroprotectants, nootropic agents, CNS modulators, antiamyloidogenics.

Some exemplary embodiments provided herein relate to combinations ofSAMe with one or more active ingredients that can be prescribed or usedfor treatment of and/or prophylaxis of a liver disorder in a subjectinclude, but are not limited to, antiviral medication such as alphainterferon, ribavirin, lamivudine, steroids, antibiotics and zincacetate.

Some exemplary embodiments provided herein relate to combinations ofSAMe with one or more active ingredients that can be prescribed or usedfor treatment of and/or prophylaxis of a cancer in a subject include,but are not limited to, chemotherapeutic agents, drug resistancemodulators, monoclonal antibodies, cytokines (e.g. interferons andinterleukins), immunocytokines, growth factors, chemoprotectants,vaccines and other biological response modifiers.

Some exemplary embodiments relate to combinations of SAMe with one ormore active ingredients that can be prescribed or used for treatment ofand/or prophylaxis of a joint or inflammatory disease/disorder in asubject include, but are not limited to, analgesics, non-steroidalanti-inflammatory drug compounds (NSAID), disease-modifyingantirheumatic drugs (DMARDs), corticosteroids, anakinra (aninterleukin-1 receptor antagonist), COX-2 inhibition, gamma-aminobutyricacid-B (GABAB) receptor agonists, such as baclofen, GABAA potentiatingdrugs, such as the benzodiazepines tumor necrosis factor(TNF)-inhibiting drugs, and other drugs that modify the immune response(immunosuppressive drugs).

Some exemplary embodiments provided herein relate to combinations ofSAMe with one or more active ingredients that can be prescribed or usedfor treatment of and/or prophylaxis of an autoimmune disease/disorder ina subject include, but are not limited to, DMARDs, corticosteroids,anakinra (an interleukin-1 receptor antagonist), TNF-inhibiting drugs,and other drugs that modify the immune response (immunosuppressivedrugs).

Some exemplary embodiments provided herein relate to combinations ofSAMe with one or more active ingredients that can be prescribed or usedfor treatment of and/or prophylaxis of a degenerative disease/disorderin a subject include, but are not limited to, NSAIDs, COX-2 inhibition,GABAB receptor agonists, such as baclofen, and GABAA potentiating drugs,such as the benzodiazepines.

Some exemplary embodiments provided herein relate to combinations ofSAMe with one or more active ingredients that can be prescribed or usedfor treatment of and/or prophylaxis of a soft tissue disease/disorder ina subject include, but are not limited to, milnacipram, pregabalin,SNRIs, NSRIs, muscle relaxers, sedatives, painkillers, and NSAIDs.

Some exemplary embodiments provided herein relate to combinations ofSAMe with one or more active ingredients that can be prescribed or usedfor treatment of and/or prophylaxis of a genetic disease/disorderrelated to hyper or hypo methylation in a subject include, but are notlimited to methionine, MTA (5′-deoxy-5′-(methylthio)adenosine) and otherSAMe metabolites.

Some exemplary embodiments provided herein relate to combinations ofSAMe with one or more active ingredients that can be prescribed or usedfor treatment of and/or prophylaxis of a gastrointestinaldisease/disorder in a subject include, but are not limited to,5-Aminosalicylic acid (5-ASA) medications, Corticosteroids (prednisone),immunomodulatory medications such as Azathioprine (Immuran),6-Mercaptopurine (6-MP), Methotrexate and Cyclosporine (Sandimmune),commonly used antibiotics such as Metronidazole (Flagyl) andCiprofloxacin (Cipro) and biologic agents such as Infliximab (Remicade).

Some exemplary embodiments provided herein relate to combinations ofSAMe with one or more active ingredients that can be prescribed or usedfor treatment of and/or prophylaxis of a cardiovascular disease/disorderin a subject include, but are not limited to, statins,angiotensin-converting enzyme (ACE) inhibitors, ASA, SAMe break downproducts such as methionine, MTA and folate, cardioprotectants,vasoprotectants, coagulation inhibitors.

Some exemplary embodiments provided herein relate to combinations ofSAMe with one or more active ingredients that can be prescribed or usedfor the treatment of and/or prophylaxis of a metabolic disease in asubject including, but not limited to sulfonylureas, biguanides,thiazolidinediones, alpha-glucosidase inhibitors, meglitinides,incretin-based therapies, and DPP-4 inhibitors.

Some exemplary embodiments provided herein relate to combinations ofSAMe with one or more active ingredients that can be prescribed or usedfor treatment of and/or prophylaxis of a disorder induced in whole or inpart by oxidative or free-radical damage including, but are not limitedto, antioxidants such as Vitamin A, Vitamin C, Vitamin E, polyphenols,flavonoids, selenium, carotenoids.

Some exemplary embodiments provided herein relate to combinations ofSAMe with one or more active ingredients that can be prescribed or usedfor treatment of and/or prophylaxis of a disorder induced in whole or inpart by damage to the central nervous system such as brain injury orspinal cord injury including, but not limited to, neuroprotectants,nootropic agents, CNS modulators, analgesics, muscle relaxants,apoptosis inhibitors, bone modulators, antioxidants.

Some exemplary embodiments provided herein relate to combinations ofSAMe with methionine, MTA, folate, vitamin B6 and/or B12. These agentsmay be correlated with lowering homocysteine production. Therefore, itis considered that combining SAMe with methionine, MTA, folate, vitaminB6 and/or B12 may result in increased supplementation of SAMe byenhancing the body's natural ability to make SAMe while at the same timesupplementing SAMe with exogenous SAMe exhibiting enhanced absorptionand improved bioavailability. As used herein the term “folate” refers tovitamin B9 in all of its natural and synthetic forms including, but notlimited to, folic acid, tetrahydrofolate and L-methylfolate.

In some embodiments, an exemplary enhanced-absorption SAMe dosage formprovided herein may be included in a kit with a separate dosage formcontaining at least one other active ingredient, exemplified by one ormore compounds suitable for the treatment of or commonly prescribed orused for the treating and/or prophylaxis in a subject a disease ordisorder selected from the group consisting of, but not limited to, amental or psychiatric disorder (e.g. psychotic/mood or non-psychoticmental disorders such as depression and substance related disorders,respectively), a nervous system disease/disorder (e.g. a central nervoussystem disease such as Alzheimer's), other neurologicaldisease/disorders (e.g. headaches and sleep disorders), conditionsassociated with injury to the central nervous system, a liverdisease/disorder (e.g. alcoholic liver disease), a cancer (e.g. solidand blood-borne cancers), a joint disease/disorder (e.g. arthritis), aninflammatory disease/disorder (e.g. ulcerative colitis), an autoimmunedisease/disorder (e.g. systemic lupus erythematosis and rheumatoidarthritis), a degenerative disease/disorder (e.g. Amyotrophic LateralSclerosis), a soft-tissue disease/disorder (e.g. a fibromyalgiadisorder), a pain disease/disorder, a genetic disorder related to hyperor hypo methylation, a gastrointestinal disease/disorder, acardiovascular disease/disorder, and a disorder induced in whole or inpart by oxidative or free-radical damage, comprising administering tosaid subject an exemplary composition provided herein which improves theabsorption of a physiologically effective amount of exogenous SAMe.

In addition to combinations of SAMe with the one or more additionalingredients exemplified above or methionine, MTA, folate, vitamin B6and/or B12, administration of the exemplary enhanced-absorption SAMeformulations provided herein may also augment the effects of other drugsor nutritional supplements being taken by the subject. Thus, someexemplary embodiments provided herein relate to combinations ofenhanced-absorption SAMe formulations with drugs or nutritionalcompounds already employed for treating other diseases for increasingthe activity of said drugs or nutritional compounds.

The present embodiments are further described by the following examples.These examples, while illustrating certain specific aspects providedherein, should not be considered to limit or circumscribe the scope ofthe disclosed embodiments.

EXAMPLES Example 1 Altered SAMe Coating Compositions Result in GISegment-Specific SAMe Absorption

In order to better understand the absorption characteristics of SAMe invivo, standard, uncoated tablets comprising SAMe were first generatedand then covered with a segment-specific coating targeting one of threedistinct regions of the GI tract.

The uncoated SAMe tablets comprising microcrystalline cellulose,croscarmellose, colloidal silicon dioxide and magnesium stearate weremade using standard procedures known to those skilled in these arts. Inorder to improve the compressibility of the composition, SAMe powder wasgranulated using a dry compaction process. Each excipient was splitbetween the intra-granular and extra-granular phases. The finaltableting mixture was compressed using a rotary tablet press fitted withelongated oval tooling at one station and the remaining stations blockedoff. The relative ambient humidity was maintained at around 30% or lessand ambient temperature was controlled between 20 and 30° C. throughoutthe process. The granules used in this formulation had good flowproperties and demonstrated no sticking or picking during compression.

In order to target SAMe release in the proximal GI tract (duodenum &jejunum) a commonly used Eudragit® coating known to dissolve above a pHof about 5.5, was applied to the SAMe tablets (EUDRAGIT is a registeredtrademark of Rohm GmbH; Darmstadt, Germany). In addition, prior toapplying the pH-dependent coating, a commercially available seal coatwas first applied.

A second formulation comprising a second commercially availableEudragit® coating was utilized to deliver SAMe to the distal GI tract(ileum/ascending colon) as it dissolves at a pH above about 7.0. Asabove, this formulation was first prepared with a commercially availableseal coat.

Finally, a rate controlling coating used to provide metered SAMe releasethroughout the entire GI tract was applied to uncoated SAMe core tabletsby a Contract Research Organization.

As seen in FIG. 1A, delivery of SAMe is achieved in all three regions ofthe GI tract and each site results in a unique pharmacokinetic profileof SAMe with different Cmax and Tmax values (FIG. 1B). In the case ofthe pH 5.5 and pH 7.0 coated tablets, the expected time of un-coating isa function of the pH sensitivity of the coating together with theexpected transit or arrival time for the targeted segments. The Tmaxobserved experimentally corresponded to that anticipated time. In thecase of the rate-controlling coating which is a non-dissolving coating,there is no seal coating; the Tmax corresponds to maximal drug deliveryin the colon.

The three formulations therefore were confirmed to show targeteddelivery at three distinct sites within the GI tract corresponding toproximal, distal and extended GI segments as judged by their Tmax. Thecore of the three products and the dose applied was identical but theamount delivered as indicated by the Cmax and the AUC was significantlydifferent. The relative bioavailability correlates with the tightjunction porosity of the targeted segment.

Example 2 In Vivo Delivery and Analysis of Absorption-Enhancing Agents

Use of absorption enhancers as a means to increase the absorption andthus bioavailability of a novel preparation of SAMe is achieved byeither co-formulating SAMe with one or more absorption enhancers orco-administering SAMe with one or more absorption-enhancing agents.Co-administration may not necessarily be at the same time as it may bemore efficacious to administer said absorption enhancers within areasonable time either before or after administration of saidproprietary preparation of SAMe.

Identification of suitable absorption enhancers may be found in the artor may be achieved in vivo. In vivo activity of compositions comprisingSAMe and one or more absorption enhancing agent may be measured afteradministration into an animal model. Preferably, the animal modelcomprises a pharmacokinetic (PK) model wherein candidate formulationsare administered using pharmacologically effective doses to non-rodentanimals (for example dog, pig, mini-pig, or primate) and blood, urine,cerebrospinal fluid (CSF) or other appropriate biological fluid isremoved at periodic intervals. The biological fluid is tested for activecompound in order to construct concentration vs. time profiles. Thesedata are analyzed and pharmacokinetic parameters are calculated in orderto assess in vivo pharmacokinetic activity. The most commonpharmacokinetic parameters analyzed in such models are Cmax, Tmax, andarea under the curve (AUC).

Alternatively, or in addition to the PK model, one can identify suitableabsorption enhancers using efficacy as a measurement, through the use ofnon-rodent models for liver disease or osteoarthritis as an example.

Plasma and urine markers include measuring markers suitable for eachdisease/disorder.

Changes in gene expression include serial analysis of gene expression(genomics) and changes in protein expression (proteomics) or changes inmetabolite levels (metabolomics).

Example 3 In Vitro Screening of Absorption-Enhancing Agents

In addition to above, identification of suitable absorption enhancersmay also be achieved using simple, standard in vitro screening assays.In the present embodiment, permeability of SAMe across Caco-2 cellmonolayers treated with an absorption enhancer is used to identifyagents which increase the amount of SAMe absorbed by said Caco-2 cellsin comparison to untreated Caco-2 cell monolayers. The Caco-2 cell lineis derived from a human colorectal carcinoma and is widely used for invitro cell culture models for the study of gastrointestinal drugabsorption (Stewart, B., (1995) Pharm. Res. 12:693). In these models,pure cell lines are grown on a semi-permeable membrane. Drugformulations are placed on the apical or basolateral side of the cellmonolayer and transport is determined via measurement of drugconcentrations on the other side of the membrane.

The Caco-2 cell line utilized here was from the American Type CultureCollection (ATCC). Caco-2 cells are grown in Dulbecco's modified Eagle'smedium (DMEM, Gibco) supplemented with 20% FBS (fetal bovine serum,Gibco), 100 uM non-essential amino acids (NEAA, Gibco) and 2 mML-glutamine (Gibco). A Beckton Dickinson BIOCOAT® HTS Caco-2 AssaySystem Kit is used resulting in 6.6×10⁵ cells/cm² seeding density(BIOCOAT is a registered trademark of Collaborative Biomedical Products,Inc., Bedford, Mass., USA). The cells used in transport studies aregrown for 3 days before the experiments. The culturing conditions are37° C. in an atmosphere of 5% CO₂ and 100% humidity.

For permeability across Caco-2 cell monolayers, the transport mediumused was Hank's Buffered Salt Solution (HBSS; purchased from Gibco)containing D-glucose, and HEPES pH adjusted to 7.4. A 2 mM aqueoussolution of either SAMe tosylate disulfate or SAMe 1,4 butanedisulfonatewas added on the apical or basolateral side according to themanufacturer's procedure for the Caco-2 kit. Samples were measured after120 minute incubation by liquid chromatography-mass spectrometry(LC/MS). The integrity of the monolayers was monitored using LuciferYellow Assay. As an example, the effect of the absorption enhancer (andspecifically a tight junction opening agent), EDTA (2 mM in wells), aswell as calcium-free medium on Caco-2 permeability of SAMe is comparedto absorption of SAMe on its own. Propranolol is a high permeabilitymarker and was utilized as a positive control for a readily absorbedmolecule.

The results in Table 2 below as well as those depicted in FIG. 2 showthat SAMe absorption on its own is low for both salts as evidenced by alow apparent permeability coefficients (Papp=0.41×10⁻⁶ and 0.50×10⁻⁶ cms⁻¹ in apical to basolateral and basolateral to apical directions,respectively for SAMe disulfate tosylate; and Papp=0.50×10⁻⁶ and0.60×10⁻⁶ cm s⁻¹ in apical to basolateral and basolateral to apicaldirections, respectively for SAMe 1,4 butanedisulfonate). Interestingly,the two stable salts of SAMe, disulfate-tosylate and1,4-butanedisulfonate, had identical permeability profiles withinexperimental error providing the evidence of their biologicalequivalence. The remaining permeability studies were carried out withSAMe disulfate-tosylate.

The measured permeability values of the SAMe salts were much lower thanthose measured with the high permeability marker, propranolol (22.4×10⁻⁶and 18.4×10⁻⁶ cms⁻1, respectively.) Permeability coefficients that areconcentration independent and/or similar in apical to basolateral aswell as basolateral to apical directions are said to be characteristicfor paracellular transport. Paracellular transport mechanism for SAMewas supported here by a 13-24 fold increase in SAMe permeability in thecalcium-free buffer as well as a 1.3-5.0 fold increase when in thepresence of a known tight junction opener, EDTA (as shown in Table 2 andFIG. 2).

TABLE 2 Permeability of SAMe Across a Monolayer of Caco-2 Cells Apicalto Basal to Basal × 10⁻⁶ cm s⁻¹ Apical × 10⁻⁶ cm s⁻¹ Well ContentsPapp_a-b Papp_a-b SD Papp_b-a Papp_b-a SD SAMe 1,4 butanedisulfonate0.50 0.08 0.60 0.13 SAMe disulfate tosylate 0.41 0.04 0.50 0.03 SAMedisulfate tosylate 9.60 0.56 6.48 0.75 Calcium-free SAMe disulfatetosylate with 0.54 0.04 2.63 0.16 EDTA Propranolol (Control) 22.44 1.7318.36 0.34

Example 3a SAMe Permeability is Increased Both In Vitro and In Vivo inthe Presence of Tight Junction Modulators

Additional Caco-2 testing was performed on a number of tight junctionopening agents as well as in vivo studies on two candidate tightjunction opening agents at a contract research organization. Similar tothe description above, the Caco-2 cell line was obtained from ATCC andgrown in DMEM (Sigma-Aldrich) supplemented with 20% FBS (Sigma-Aldrich),100 uM non-essential amino acids (Sigma-Aldrich) and 2 mM L-glutamine(Sigma-Aldrich). The Caco-2 cells grown in tissue culture flasks weretrypsinized, suspended in medium, and the suspensions were applied towells of a collagen-coated BioCoat Cell Environment in 24-well format at24,500 cells per well. The cells were allowed to grow and differentiatefor three weeks, feeding at 2-day intervals.

For apical to basolateral permeability, a 2 mM aqueous solution of SAMetosylate disulfate was added to the apical side and the amount ofpermeation was determined on the basolateral side. The apical andbasolateral side buffers contained modified Transport Buffer (25 mMHEPES, 1× Hank's Balanced Salt Solution (Sigm-Aldrich)) pH 7.4. Caco-2cells were incubated with these buffers for 2 hours, and the receiverside buffer was removed for analysis by LC/MS/MS. The receiver, donor,and dosing solution were diluted with an equal volume of 0.2 N HClimmediately after the assay in order to increase SAMe stability. Donorand dosing solution were diluted 100-fold to ensure that theconcentration was within the linear range of the assay.

To confirm the integrity of the Caco-2 cell monolayers, TEER (TransEpithelial Electrical Resistance) measurements were performed on eachwell at the end of the experiment.

The permeability (Papp) of SAMe is calculated using the followingformula:

${Papp} = \frac{Q}{\frac{t}{CoA}}$

Where dQ/dt is the rate of permeation, C₀ is the initial concentrationof test agent, and A is the area of the monolayer.

As shown in Table 3 below, the presence of two different fatty acids,either a C10 fatty acid or a sulfonic acid, resulted in a dramaticincrease in the permeability of SAMe.

The zwitterionic surfactants,3-(N,N-Dimethylpalmitylammonio)propanesulfonate and palmitoyl carnitinechloride were also each tested for their effect on SAMe uptake and bothshowed marked increase in SAMe permeability as seen in Table 3. Inaddition, alpha-cyclodextrin and a dicarboxcylic acid each resulted in a5-6 fold increase in SAMe permeability. The low and high permeabilitycontrols, ranitidine and warfarin, respectively, verify the use of theseabsorption enhancing agents as an in vitro screening method formeasuring the effect of these agents on SAMe permeability across Caco-2monolayers. Furthermore, the permeability of SAMe alone, versus SAMe inthe absence of calcium or in the presence of the tight junctionmodulators, palmitoyl carnitine chloride or caprate is depicted in thegraph in FIG. 3A.

TABLE 3 Permeability of SAMe in the Presence of Various Tight JunctionModulators Mean Permeability TEER Fold Chemical Caco-2 CoefficientResistance Transport Test Article Class Concentration (×10⁻⁶ cm/s)(ohms) Increase SAMe API control 2 mM 0.14 (run 2) 782 1X   (control)1.6 (run 3) 901 1X   1.6 (run 4) 1004 1X   Sodium Caprate Fatty acid 14mM 1.79 (run 2) 235 12.8X  9.1 (run 4) 260 5.7X 7 mM 4.1 (run 4) 9052.6X Sodium 1- Fatty acid 14 mM 3.1 (run 2) 120 21.9X  decanesulfonate15.5 (run 3) 135 9.7X 7 mM 6.3 (run 3) 165 3.9X PalmityldimethylZwitterionic 0.6 mM 3.8 (run 4) 415 2.4X ammonio propane surfactant 0.3mM 6.0 (run 3) 420 3.8X sulfonate 1.4 (run 4) 434 0.9X PalmitoylZwitterionic 0.15 mM 0.78 (run 2) 272 5.6X Carnitine surfactant 5.0 (run4) 519 3.1X Chloride (PCC) 0.4 mM 10.0 (run 4) 366 6.3X DodecyltrimethylFatty amine 14 mM 10.5 (run 4) 431 6.6X Ammonium Bromide Alpha-Cyclodextrin 51.4 mM 0.75 (run 2) 293 5.4X Cyclodextrin 6.3 (run 4) 3413.9X Dicarboxylic acid Organic acid 66.6 mM 10.5 (run 3) 143 6.6X 15.5(run 4) 136 9.7X Ranitidine Low 50 uM 0.91 (run 2) 1080 n/a permeability0.91 (run 3) 975 n/a control 1.0 (run 4) 931 n/a Warfarin High 50 uM48.8 (run 2) 976 n/a permeability 49.0 (run 3) 941 n/a control 46.6 (run4) 1011 n/a

To further investigate the effects of these tight junction modulators onSAMe absorption, in vivo pharmacokinetic testing of caprate and PCC wereperformed using animal subjects. Caprate is one of the mostwell-recognized tight junction modulators used in pharmaceuticaldevelopment. Formulations comprising caprate have advanced through toPhase III clinical trials demonstrating its effectiveness and safety.For this reason, formulations comprising SAMe as well as caprate werefirst investigated for pharmacokinetic analysis in vivo.

Fasted male beagle dogs (7-10 kg) were used to conduct a single dose,crossover study to compare the bioavailability of SAMe+caprate enhancerwith SAMe control capsules (Table 4). The study protocol was approved bythe institution's Animal Care Committee, and all animals were cared foraccording to regulations proposed by Agriculture Canada and the USDA.Each group (6 dogs for SAMe/caprate and 6 dogs for the control) wasdosed with a single orally administered enteric coated capsule, underfasted conditions, followed by 5 mL of purified water orally with asyringe to facilitate swallowing. Pharmacokinetic blood samples (2 mLeach) for SAMe analysis were collected from the jugular vein using thefollowing timepoints: pre-dose, 20 and 40 minutes, 1, 1.5, 2, 3, 4, 6,and 8 hours after treatment. The venipuncture blood samples werecollected into tubes containing the anticoagulant K2-EDTA, and stored onwet ice pending processing. Following collection, samples werecentrifuged (at 4° C.) to separate the plasma fraction from the bloodcells. The resulting plasma fraction was recovered and stored frozen (at−80° C.) using polypropylene tubes pending bioanalytical analysis.

The concentration of SAMe in dog plasma was determined using a wellestablished LC/MS/MS method. This method employs stable-isotope dilutionliquid chromatography-electrospray injection tandem mass spectrometry(LC-ESI-MS/MS) to determine SAMe and SAH in plasma. The analysis used tocalculate the main pharmacokinetic parameters (Cmax, T_(max), and AUC)was conducted using GraphPad Prism® 5 software.

The capsules tested in beagle dogs were formulated using size #13gelatin capsules, each containing 200 mg SAMe ion. The capsulescontained varying amounts of additional excipients (Microcrystallinecellulose, Sodium Starch Glycolate, Colloidal Silicon Dioxide, andMagnesium Stearate), with and without 200 mg of caprate enhancer.Capsules were manually filled and the resulting formulations aresummarized in Table 4:

TABLE 4 Capsule formulations of SAMe with and without Sodium CaprateControl capsule Caprate capsule Component (mg/capsule) (mg/capsule)Capsule Core SAMe disulfate tosylate 384.5 384.5 MicrocrystallineCellulose 113 79.3 79.3 Microcrystalline Cellulose 112 803.1 803.1Sodium Starch Glycolate Type A 27.5 27.5 Colloidal Silicon Dioxide 2.82.8 Magnesium Stearate 5.5 5.5 Sodium Caprate 0 200 (representing 176 mgof caprate)

Prior to applying an enteric coating to the capsules, a seal coat wasfirst applied in order to improve the capsule surface properties. Astirred 12% suspension of a commercially available seal coat in purifiedwater was applied to the uncoated SAMe capsules in an Aeromatic CoatingColumn using 55° C. inlet air temperature and 4-6 g/min spray rate until4% weight gain was achieved (Table 5). A plasticized 80% solid (w/w)commercially available enteric coat designed to dissolve at pH 5.5 wasthen applied to the tablet formulations such that they would remainintact within the stomach. The enteric coating was generated using astirred 30% aqueous suspension (56% of final coating suspension byweight), plasticizer 20% aqueous suspension (7% of final coatingsuspension by weight), triethyl citrate (2% of final coating suspensionby weight) and purified water (35% of final coating suspension byweight). These components were mixed and then applied to the seal coatedSAMe capsules in an Aeromatic Coating Column using 55° C. inlet airtemperature and 4-6 g/min spray rate, until a 6% weight gain wasachieved (Table 5).

TABLE 5 Coating composition for capsules Seal Coat Control capsuleCaprate capsule Composition (% weight (% weight Component (% of Solids)of capsule) of capsule) Seal Coat LustreClear LC103  93% 4% 4% FerricOxide 0.7% Titanium Dioxide 3.3% Simethicone 2.8% Enteric Coat EudragitL30 D55 86.8%  6% 6% Triethyl Citrate 10.3%  Plasacryl T20 2.9%

Surprisingly and quite unexpectedly, the presence of caprate did notenhance the in vivo bioavailability of SAMe as shown in FIG. 3B. Thecontrol Cmax was 1627 ng/mL compared to a Cmax of 1597 ng/mL for thecaprate formulations and the AUC values were 4191 hg hr/mL (control)versus 4510 ng hr/mL (with caprate). Even more surprising is the data inFIG. 3C, which shows the effect of the zwitterionic surfactant,palmitoylcarnitine chloride (PCC), on SAMe bioavailability. Experimentswere carried out as described above for caprate except that PCC wassubstituted for formulations in place of caprate.

Capsules comprising SAMe and 200 mg of DL-palmitoylcarnitine chloride(representing 183.4 mg of the DL-palmitoylcarnitine ion) wereadministered to 6 male beagle dogs as above and the amount of SAMe inthe plasma was compared to the control group of 6 male beagle dogsadministered a single dose of the control (SAMe alone). The compositionof the PCC formulation is set forth in Table 6.

TABLE 6 Capsule formulations of SAMe with and without PCC Controlcapsule PCC capsule Component (mg/capsule) (mg/capsule) Capsule CoreSAMe disulfate tosylate 384.5 384.5 Microcrystalline Cellulose 113 79.379.3 Microcrystalline Cellulose 112 803.1 803.1 Sodium Starch GlycolateType A 27.5 27.5 Colloidal Silicon Dioxide 2.8 2.8 Magnesium Stearate5.5 5.5 DL-palmitoylcarnitine chloride (PCC) 0 200 (representing 183.4mg of DL-palmitoyl- carnitine ion)

Surprisingly, the presence of PCC resulted in a significant increase inthe amount of SAMe present in the plasma as compared to the controlsubjects (FIG. 3C). The SAMe Cmax in the presence of PCC wasapproximately 91% higher than the Cmax of the control group (3108 ng/mLand 1627 ng/mL, respectively). In addition, the SAMe AUC increased byapproximately 86% in the presence of PCC (7794 ng hr/mL compared to 4191ng hr/mL for the control).

This data, together with the in vitro screening results in Table 3above, show for the first time that a subset of high potency tightjunction opening absorption enhancers exists and that identification ofpotential high potency tight junction opening agents may be carried outusing, for example, a Caco-2 cell model as described above. Based on theconcentration of tight junction modulator utilized as well as theincrease in SAMe permeability achieved, suitable high potency tightjunction modulators elicit at least a two-fold increase in SAMe uptakein vitro when used at low concentrations (i.e. about 2 mM or below).More preferably, at least a three-fold increase in SAMe uptake occurs atlow concentrations of these tight junction opening agents and, even morepreferably, a four-fold or greater increase is achieved.

Example 3b The Effect of Additional Zwitterionic Surfactants on SAMePermeability and Bioavailability

In vitro and in vivo experiments are carried out as described in Example3a using acyl carnitines with either longer or shorter chain length incomparison to PCC (see previous Table detailing acyl carnitines). Forexample, Caco-2 cell and beagle experiments as described above areconducted using stearoyl carnitine, myristoyl carnitine, lauroylcarnitine, decanoyl carnitine or mixtures of two or more thereof.

Experiments described here and above may also be carried out using othersolid or semi-solid, oral dosage forms, such as tablets. Tablets aretypically generated as described in Example 1 as well as in co-owned andco-pending U.S. patent application Ser. No. 12/845,600.

Example 3c SAMe Permeability and Bioavailability Increase in thePresence of Various Sulfobetaines

In vitro and in vivo experiments measuring SAMe permeability andpharmacokinetic parameters, respectively, as described in Example 3awere performed using dimethylammoniopropane sulfonates sulfobetaines) ofvarying chain length (carbon chain length of 10-18 as listed below).

In vitro Caco-2 testing was performed as detailed above using 0.15 mM ofsulfobetaine-10, sulfobetaine-12, sulfobetaine-14, sulfobetaine-16, orsulfobetaine-18 as the enhancer. The presence of sulfobetaine-10 orsulfobetaine-12 did not improve SAMe permeability across the Caco-2 cellmonolayer. However, as shown in Table 7 below, the presence ofsulfobetaine-14, sulfobetaine-16 or sulfobetaine-18 at a concentrationof 0.15 mM resulted in at least a five-fold increase in SAMepermeability across the Caco-2 cell monolayer, and permeabilityincreased with increasing acyl chain length. These three sulfobetaineswere thus identified as high potency absorption enhancers as defined inthe present application.

As part of these in vitro studies, the degree of Caco-2 cell recovery(as observed by degree of permeability and cell attachment 24 hours posttreatment with each tight junction modulator) was also investigated.This 24 hour resistance recovery test is an indirect measure ofepithelial cell disruption and data supporting a trend toward less invitro disruption may be associated with less impact in vivo. The resultsin Table 7 show that cell recovery improves with decreasing sulfobetainechain length.

TABLE 7 Permeability of SAMe in the Presence of Various SulfobetainesTEER Mean Resist. Permeability Fold TEER 24 hour Caco-2 CoefficientTransport Resistance post assay Enhancer Conc. (×10⁻⁶ cm/s) Increase(ohms) (ohms) SAMe   2 mM 9.6 1X 462 control 0.14 1X 782 1.6 1X 901 1.61X 1004 0.14 1X 1045 2280 0.17 1X 1031 2181 0.26 1X 726 1440 SAMe   2 mM0.186 1X 924 1975 control 0.131 1X 1285 1871 0.124 1X 995 1740 Sulfobe-0.15 mM 0.218   1.2X 1043 1553 taine-8 Sulfobe- 0.15 mM 0.257   1.4X 9341923 taine-12 Sulfobe- 0.15 mM 0.654   5.0X 263 1892 taine-14 Sulfobe- 0.6 mM 3.8   2.4X 415 taine-16 (Palmityl- 7.4  51.9X 25 33 dimethyl 0.3 mM 6.0   3.8X 420 ammonio 1.4   0.9X 434 propane) 2.5  17.7X 41 84sulfonate  0.3 mM 2.19  17.7X 71 30 0.15 mM 1.9  13.3X 99 1602 0.678  5.2X 123 292 Sulfobe- 0.15 mM 0.649   5.0X 123 292 taine-18

To further analyze the effects of these tight junction modulators, invivo experiments using fasted male Beagle dogs as detailed in Example 3awere carried out using SAMe capsules comprising either sulfobetaine-14or sulfobetaine-16. Capsules comprising 200 mg SAMe (ion) and 200 mg ofsulfobetaine-14 or sulfobetaine-16 were administered to six male beagledogs as above and the amount of SAMe in the plasma was compared to thecontrol group of six male beagle dogs administered a single dose of thecontrol (200 SAMe (ion) alone). The compositions of the sulfobetaine andcontrol formulations are set forth below in Table 8.

TABLE 8 Capsule formulations of SAMe with and without Sulfobetaine-14 orSulfobetaine-16 Control capsule Sulfobetaine capsule Component(mg/capsule) (mg/capsule) Capsule Core SAMe disulfate tosylate 384.5¹384.5¹ Microcrystalline Cellulose 113 79.3 79.3 MicrocrystallineCellulose 112 803.1 803.1 Sodium Starch Glycolate Type A 27.5 27.5Colloidal Silicon Dioxide 2.8 2.8 Magnesium Stearate 5.5 5.5Sulfobetaine-14 or Sulfobetaine-16 0 200

As identified in FIG. 4A, the presence of sulfobetaine-16 resulted in asignificant increase in the amount of SAMe present in the plasma ascompared to the control subjects. As shown, the SAMe Cmax in thepresence of sulfobetaine-16 was approximately 316% higher than the Cmaxof the control group. In addition, the SAMe AUC increased byapproximately 313% in the presence of sulfobetaine-16. Surprisingly, thepresence of sulfobetaine-14 also resulted in a significant increase inthe bioavailability of SAMe as measured by an increase in the averageSAMe Cmax from 1627 ng/mL for the control to 4148 ng/mL in the presenceof sulfobetaine-14 (FIG. 4B). The average SAMe AUC also increased from4191 ng·hr/mL (control group) to 10226 ng·hr/mL in the presence ofsulfobetaine-14.

This data supports the finding that a subset of high potency tightjunction opening absorption enhancers exists and that identification ofsuitable high potency tight junction opening agents for increasing SAMebioavailability in vivo may be carried out using, for example, a Caco-2cell model as described above.

Example 4 Plasma Levels of SAMe Metabolites are not Elevated Relative toPlasma Levels of SAMe

The data presented above clearly demonstrates for the first time thatthe use of absorption enhancers to increase SAMe permeability in vivoleads to enhanced SAMe bioavailability. This finding is in contrast tothe general dogma surrounding SAMe metabolism which continually reportsthat low SAMe bioavailability is due to active metabolism by the liverupon administration. If this were the case it would be reasonable tobelieve that the plasma level of one or more SAMe metabolites would besignificantly elevated after administration. In order to test thistheory, the inventors measured the level of S-adenosyl homocysteine(SAH), the primary metabolite of SAMe, at various time points afteradministration of a 1600 mg dose of a commercially available SAMeformulation.

As seen in FIG. 5, at all time points measured, the plasma concentrationof SAH (which is plotted on a scale that is tenfold lower than the SAMeconcentration) is significantly lower than the level of SAMe itself.Additional SAMe metabolites were also measured and, similarly to SAH,there were no differences in their baseline levels (results not shown.)

These results demonstrate that SAMe metabolism is not responsible forthe low bioavailability of SAMe upon administration.

Example 5 SAMe Delivered to the Stomach can be Absorbed into the Plasma

Those skilled in these arts are also of the view that SAMe delivery mustbypass the stomach in order to avoid its harsh low-pH environment whichis taught to cause degradation. In order to even further understand themechanism of SAMe absorption in vivo, and thus better control itsbioavailability, the release of SAMe into the stomach environment andits absorption there from was also assessed.

Uncoated SAMe was formulated into tablets comprising microcrystallinecellulose, croscarmellose, colloidal silicon dioxide and magnesiumstearate using standard procedures as described in Example 1.

The absence of an enteric coating in this formulation was intended tocause release of SAMe within the stomach. The resulting pharmacokineticprofile was studied by measuring the presence of the drug in plasma atvarious time points after administration. A single dose of 400 mg SAMewas given to seven healthy and fasted, male volunteers.

As seen in FIG. 6, the average Cmax of the seven subjects administeredthe uncoated SAMe formulation was about 145 ng/mL for the 400 mg dose.These results show that, contrary to what is repeatedly reported in theart, SAMe can be delivered to the stomach without using enteric coatedformulations and still give rise to increased absorption as seen withthe plasma SAMe levels reported here.

1.-103. (canceled)
 104. A pharmaceutical composition comprising atherapeutically effective dosage of S-adenosylmethionine and at leastone zwitterionic surfactant.
 105. The composition of claim 104, whereinsaid zwitterionic surfactant enhances systemic delivery ofS-adenosylmethionine as measured by an increase in C_(max) and/or AUCcompared to a control composition that lacks said zwitterionicsurfactant.
 106. The composition of claim 105, wherein the C_(max)and/or AUC of said composition is at least 140% of the C_(max) and/orAUC obtained with a control composition that lacks said zwitterionicsurfactant.
 107. The composition of claim 104, wherein said compositionis a non-parenteral composition.
 108. The composition of claim 107,wherein the non-parenteral composition is an oral dosage composition.109. The composition of claim 108, wherein the oral dosage compositionis formulated as a tablet, a capsule, or a multiparticulate formulation.110. The composition of claim 108, wherein the oral dosage compositioncomprises about 50 to about 500 mg of S-adenosylmethionine.
 111. Thecomposition of claim 107, wherein the non-parenteral composition isformulated as a dietary supplement or a medical food.
 112. Thecomposition of claim 107, wherein at least a portion of thenon-parenteral composition is formulated to dissolve in at least one ofthe stomach, duodenum, jejunum, ileum, large intestine, or colon. 113.The composition of claim 107, wherein the non-parenteral compositioncomprises a pH sensitive coating.
 114. The composition of claim 104,wherein said zwitterionic surfactant is an acyl carnitine.
 115. Thecomposition of claim 114, wherein said acyl carnitine is a palmitoylcarnitine, lauroyl carnitine, stearoyl carnitine, myristoyl carnitine,decanoyl carnitine, or a salt thereof.
 116. The composition of claim104, wherein said zwitterionic surfactant is a sulfobetaine.
 117. Thecomposition of claim 116, wherein said sulfobetaine is sulfobetaine-10,sulfobetaine-12, sulfobetaine-14, sulfobetaine-16, or sulfobetaine-18.118. The composition of claim 104, wherein said zwitterionic surfactantis a high potency absorption enhancer as determined in a Caco-2monolayer permeability assay.
 119. The composition of claim 104, whereinS-adenosylmethionine and said zwitterionic surfactant are in a ratio offrom 1:100,000 to 100,000:1; 1:150 to 150:1; or 1:10 to 10:1(weight:weight).
 120. The composition of claim 104, whereinS-adenosylmethionine and said zwitterionic surfactant are in a ratio of1:1 (weight:weight).
 121. The composition of claim 104, wherein thecomposition comprises 50 to 3200 mg of S-adenosylmethionine.
 122. Thecomposition of claim 104, further comprising at least one delayedrelease component.
 123. The composition of claim 122, wherein thedelayed release component is a pH-triggered enteric coating.
 124. Thecomposition of claim 122, wherein the delayed release component isformulated to dissolve in at least one of the duodenum, jejunum, ileum,large intestine, or colon.
 125. A method for increasing thebioavailability of exogenous SAMe administered to a subject, said methodcomprising administering to the subject a non-parenteral compositioncomprising a therapeutically effective dosage of the composition ofclaim
 104. 126. The method of claim 125, wherein the composition is anoral dosage composition.
 127. The method of claim 125, wherein thecomposition is formulated as a dietary supplement or a medical food.128. The method of claim 125, wherein at least a portion of thecomposition is formulated to dissolve in at least one of the stomach,duodenum, jejunum, ileum, large intestine, or colon.
 129. The method ofclaim 128, wherein the composition comprises a pH sensitive coating.130. The method of claim 125, wherein said zwitterionic surfactant isadministered either before or after administration of the compositioncomprising the at least one therapeutically effective dosage ofS-adenosylmethionine.
 131. A method of treating a disorder selected fromthe group consisting of a mental or psychiatric disorder, a nervoussystem disease or disorder, a neurological disease or disorders, acondition associated with injury to the central nervous system, a liverdisease or disorder, a cancer, a joint disease or disorder, aninflammatory disease or disorder, an autoimmune disease or disorder, adegenerative disease or disorder, a soft-tissue disease or disorder, apain disease or disorder, a genetic disorder related to hyper- orhypo-methylation, a gastrointestinal disease or disorder, acardiovascular disease or disorder, and a disorder induced in whole orin part by oxidative or free-radical damage, comprising administering toa patient in need thereof a composition of claim
 104. 132. The method ofclaim 131, wherein: (i) the mental or psychiatric disorder is selectedfrom the group consisting of an anxiety disorder, schizophrenia, majordepressive disorder, multi-infarct dementia, minor depression,postpartum depression, inflammatory depression, late-life depression,Parkinson's depression, HIV-associated depression, and bipolar disorder;(ii) the inflammatory disease or disorder is selected from the groupconsisting of systemic lupus, inflammatory bowel disease, allergicrhinitis, contact dermatitis, asthma, autoimmune hepatitis, and pelvicinflammatory disease; (iii) the cardiovascular disease or disorder isselected from the group consisting of hyper- or hypo-homocysteinemia,coronary heart disease, stroke, peripheral vascular disease, andatherosclerotic disease; (iv) the depressive disorder is a comorbiddepression arising in a subject who is or has been undergoing treatmentfor one or more diseases or disorders selected from the group consistingof cancer, Parkinson's and HIV; (v) the nervous system disease ordisorder or injury is selected from the group consisting of Parkinson'sdisease, Alzheimer's disease, and cognitive impairment; (vi) the liverdisease or disorder is selected from the group consisting of alcoholicliver disease, non-alcoholic fatty liver disease, viral or non-viralhepatitis, liver cancer, oxidative liver disease, drug induced liverinjury, cholestasis, and cirrhosis; (vii) the cancer is selected fromthe group consisting of liver cancer, colon cancer, rectal cancer,stomach cancer, esophageal cancer, and adenocarcinoma; (viii) the jointdisease or disorder is arthritis or osteoarthritis; (ix) the soft-tissuedisease or disorder is fibromyalgia; (x) the pain disease or disorder isselected from the group consisting of fibromyalgia, and abdominal pain;or (xi) the genetic disorder related to hyper- or hypo-methylation ismethylenetetrahydrofolate reductase deficiency.